PHYSIOLOGY OF THE ENDOCRINE SYSTEM

General notions about the endocrine system

Endocrine system together with nervous system are the systems which are involved
in regulation and coordination of the activity of all cells in the body. The action of the nervous
system is rapid, immediate and of short duration. Usually it is localized. By comparison with
this, the activity of the endocrine system is more diffused, delayed and has a longer duration
of action. Nevertheless a clear separation between these two systems cannot be done,
because there are multiple interactions between them. Nervous system, by means of
hypothalamus, regulates the activity of the majority of endocrine glands, while many
hormones have effects on the development and function of the nervous system.
Endocrine system is made up of endocrine glands, which compare to exocrine glands,
have the characteristics that follow:
 They do not have excretory channel
 Their secretion is released directly into the blood stream
 One of the pole (basal) of the gland cells is oriented to a capillary, from where come
substances necessary for the hormone synthesis, while the apical pole is oriented
towards the venous segment of a capillary, where the hormone is released
 The secretion is represented by hormones
 Hormones usually act at a distance from the cells where they have been synthesized
 Hormones act on those cells who have receptors for them

Hormones (hormao = to stimulate) are defined as chemical messengers synthesized

by endocrine glands and transported by blood to target cells or organs. They perform several
effects:

 Influence metabolic reactions

 Are involved in storage and catabolism of carbohydrates, lipids and proteins
 Stimulate physical, mental and sexual development of the body
 Participate to autonomic reactions (adrenaline synthesized by adrenal gland influences
the autonomic activity of different organs)
 Regulates the synthesis of erythropoietin
 Control the integration of circulation with digestion and absorption of food. They are
similar in their action with enzymes.

From chemically point of view hormones can be: peptides, amino acids (derivatives
of tyrosine), proteins or steroids (lipids- derivatives of cholesterol). Many hormones
(proteins, polypeptides) are synthesised as preprohormones, but they are cleaved to
prohormone and packed in vesicles at the level of endoplasmic reticulum and Golgi apparatus.
In this process, enzymes in the vesicles cleave the prohormones to produce smaller,
biologically active hormones and inactive fragments. The vesicles are stored within the
cytoplasm, and many are bound to the cell membrane until their secretion is needed.
Steroid hormones derive from cholesterol. Although there is usually very little
hormone storage in steroid-producing endocrine cells, large stores of cholesterol esters in

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cytoplasm vacuoles can be rapidly mobilized for steroid synthesis after a stimulus. Much of
the cholesterol in steroid-producing cells comes from the plasma, but there is also de novo
synthesis of cholesterol in steroid-producing cells. Because the steroids are highly lipid
soluble, once they are synthesized, they diffuse across the cell membrane and enter the
interstitial fluid and then the blood.

Mechanism of action of hormones:

 The action is exerted only after the hormone is bound by a specific receptor.
 The number of receptors for hormone on a cell is variable, from 2,000 until 100,000
receptors per cell.
 When the concentration of a hormone in blood increases, the number of receptors
expressed at the level of a cell decreases; this is named down-regulation.
 When the concentration in blood of a hormone decreases, the number of receptors
expressed at the level of the cell increases; this is named up-regulation.
 According to the chemical nature of hormones, the receptors are found at the level of
cell membrane, cytoplasma or nucleus of the cell. Hormones that are hydrosoluble
(proteins, peptides) have the receptors are at the level of cell membrane; liposoluble
hormones (like steroid hormones and thyroid hormones) have the receptor inside of
the cell.
 They are released by exocytosis when the Ca2+ or cAMP concentration inside of cell
increases.
 A hormone can performe several effects on several organs.
 A target organ can be influenced by several hormones.
 An endocrine gland can synthesize several hormones.

Classification of hormones after their mechanism of action

Hormones together with neurotransmitters are named primary messengers. The

effect of hormones will be due to some secondary messengers found inside of the cells (Ca2+,
DAG, cAMP, IP3). Related to their mechanism of action, hormones are classified into several
categories.

A) Hormones that act by means of membrane receptors (proteins, peptides, catecholamines)

a) Receptors that use G proteins

1) Hormones which use cAMP as secondary messenger

Hormones which are polypeptides cannot penetrate inside of cells, so they have the
receptors at the level of cell membrane. After the hormone is bound by its receptor, an
enzyme is activated, named adenyl cyclase (or adenylyl cyclase) and cyclic AMP is produced.
The connection between the receptor and adenylyl cyclase is made by a protein named
protein G or (heterotrimeric GTP binding protein). Proteing G is made up of 3 subunits: alpha,
beta and gamma. It is considered that there are several types of G proteins; functionally, they
are stimulating G protein, or inhibitory G proteins: Gs or Gi.

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 When protein G is inactive, it is coupled with GDP (alpha subunit binds it). When it
becomes activated, GDP is replaced by GTP. This is due to the interaction of the hormone with
its receptor. A complex made up of hormone and its receptor can activate several molecules
of protein G. After GDP is replaced by GTP, alpha subunit detaches from beta and gamma, and
it activates adenylyl cyclase. Alpha subunit has a GTP-ase activity and hydrolysis GTP to GDP.
Adenylyl cyclase will convert ATP to cAMP. Cyclic AMP is the secondary messenger that in its
turn will activate another protein, namely protein kinase A. Protein kinase A will
phosphorylate other proteins and enzymes inside of cells and by this the effect of hormone is
produced. cAMP is broken down by the enzyme: phosphodiesterase, which converts cAMP
into 5-AMP. Phosphodiesterase can be inhibited by different substances: Coffee, Teophiline,
Methylxantine.

Mechanism of activation of a G protein–coupled receptor (after A.C. Guyton, J. Hall, Textbook

of Medical Physiology, 2006)

Several hormones performe their action by means of cAMP : ADH (acting on

receptor V2), parathyroid hormone (PTH), glucagon, adrenaline and noradrenaline (on beta
receptors), thyroid stimulating hormone (TSH), adrenocorticotrope hormone (ACTH),
luteinizing hormone (LH), folliculo stimulating hormone (FSH), angiotensin II, calcitonin.

2) Hormones that use Inositol Triphosphate (IP3) and Diacyl Glycerol (DAG) as secondary
messengers

Other hormones act by activation of another enzyme - Phospholipase C (PLC). After

binding of hormone by the specific receptor, PLC is activated. After activation, PLC breaks
down phosphatidylinositol 4,5 diphosphate (PIP2) from the membrane into inositol 1,4,5
triphosphate (IP3) and diacyl glycerol (DAG). The activation of PLC is mediated by a G protein,
too (Gq). Inositol triphosphate diffuses inside of endoplasmic reticulum and opens calcium
channels by which calcium will diffuse outside. Calcium together with a protein named
calmodulin will form a complex (4 Ca2+ for one calmodulin molecule) that will interact with
different enzymes activating them. Diacyl glycerol (DAG) will activate protein kinase C and this

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protein will phosphorylate other proteins and enzymes. By this process hormonal effect is
carried out.
There are some hormones that performe their action in this way: oxytocin,
cathecolamines (on alpha receptors), antidiuretic hormone (on V1 receptors), cholecystokinin,
histamin (on H1 receptors).

DAG and IP3 as second messengers (after A. Despopoulos, S. Silbernagl, Color Atlas of
Physiology, 2003)

b) Ion channel - linked receptors

Neurotransmitters as acetylcholine and norepinephrine, combine with receptors in

the postsynaptic membrane. That is followed by a conformational change of receptor and
opening or closing of a ion channel, that is associated with the receptor. There are channels
for Na+, Ca2+, K+ in the structure of these receptors. For instance, in a neuromuscular junction,
when an impulse reaches the terminal knob of the motor neuron, Acetylcholine is released
from the vesicles and interacts with the postsynaptic receptor (nicotinic R). This is followed by
opening of ligand gated Na+ channels and penetration of sodium inside the muscle fiber.
Sodium alters the RMP, so that endplate potential is generated.

c) Enzyme-Linked Hormone Receptors

There are receptors that function directly as enzymes or are associated with enzymes.
They have their hormone-binding site on the outside of the cell membrane and their catalytic
or enzyme-binding site on the inside. When the hormone binds to the receptor, an enzyme
inside the cell membrane is activated (or inactivated). Some receptors have intrinsic enzyme
activity, others act through enzymes that are closely associated with the receptor to produce
changes in cell function.

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  Receptors that contain tyrosine - kinase – for insulin, insulin-like growth factor
(IGF-1), epidermic growth factor, platelet derived growth factor. When the hormone binds to
the receptor, tyrosine - kinase is activated, which has two effects: 1) kinase phosphorilates the
tyrosine residues of receptor and this self phosphorilation intensifies the signal, 2) kinase can
phosphorilate the tyrosine residues from other intracellular proteins, including enzymes and
this may regulate their activity.

 Receptors that are associated with tyrosine-kinase – for Growth hormone, PRL,
erythropoietin, leptin (a hormone secreted by fat cells).

The leptin receptor (after A.C. Guyton, J. Hall, Textbook of Medical Physiology, 2006)

For the leptin receptor, one of the signaling pathways occurs through a tyrosine kinase
of the janus kinase (JAK) family, JAK2. The leptin receptor exists as a dimer, and binding of
leptin to the extracellular part of the receptor alters its conformation, enabling
phosphorylation and activation of the intracellular associated JAK2 molecules. The activated
JAK2 molecules then phosphorylate other tyrosine residues within the leptin receptor– JAK2
complex to mediate intracellular signaling. The intracellular signals include phosphorylation of
signal transducer and activator of transcription (STAT) proteins, which activates transcription
by leptin target genes to initiate protein synthesis. Phosphorylation of JAK2 also leads to
activation of other intracellular enzyme pathways such as mitogen-activated protein kinases
(MAPK) and phosphatidylinositol 3-kinase (PI3K). Some of the effects of leptin occur rapidly as
a result of activation of these intracellular enzymes, whereas other actions occur more slowly
and require synthesis of new proteins.

B) Intracellular Hormone Receptors (the receptor is in the cytoplasm/nucleus)

Other hormones - adrenal and gonadal steroid hormones, thyroid hormones, and
vitamin D, bind with protein receptors inside the cell. Because these hormones are lipid
soluble, they easily cross the cell membrane and interact with receptors in the cytoplasm or
nucleus. The activated hormone receptor complex then binds with a specific regulatory

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(promoter) sequence of the DNA called the hormone response element, and in this manner
either activates or represses transcription of specific genes and formation of messenger RNA
(mRNA). Minutes, hours, or even days after the hormone has entered the cell, newly formed
proteins appear in the cell and become the controllers of new or altered cellular functions.
The thyroid hormones (thyroxine and triiodothyronine) cause increased transcription
by specific genes in the nucleus. These hormones bind directly with receptor proteins in the
nucleus itself; these receptors are probably protein molecules located within the
chromosomal complex, and they likely control the function of the genetic promoters or
operators. They activate the genetic mechanisms for the formation of many types of
intracellular proteins ( more than 100). Once bound to the intranuclear receptors, the thyroid
hormones can continue to express their control functions for days or even weeks.

PITUITARY GLAND (HYPOPHYSIS)

It is a very small gland, having the weight of 0.5-1 g, placed at the level of sella turcica
(sphenoid bone). It is made up of 2 components: neurohypophysis (posterior pituitary) and
adenohypophysis (anterior pituitary). It is connected to the hypothalamus by
pituitary/hypophyseal stalk.
Neurohypophysis derives from diencephalon, while adenohypophysis derives from
Rathke’s pouch (pharyngeal epithelium). Both have anatomical and functional connections
with hypothalamus.
Adenohypophysis is made of pars tuberalis and pars distalis. Pars distalis is the
anterior lobe. Neurohypophysis is made up of median eminence of hypothalamus, infudibular
stem and infundibular process/pars nervosa or posterior lobe. Besides the anterior and
posterior lobes, there is also an intermediate lobe (pars intermedia), which is very small and
not very well developed in case of human beings, but very well developed in other inferior
animals.
Pituitary gland secretes several hormones : Growth hormone (GH) (or somatotropic
hormone), prolactin, thyroid stimulating hormone (TSH) (or thyrotropin), FSH (follicle
stimulating hormone), LH (Luteinizing hormone), adrenocorticotrope hormone (ACTH) ,
melanocytes stimulating hormone (MSH), ADH (antidiuretic hormone, also called
Vasopressin), and oxytocin. The activity of pituitary gland is under the controle of
hypothalamus.
The synthesis and secretion of hormones of adenohypophysis is under the control of
hypothalamus, that secretes releasing hormones (Liberins) and Inhibitory hormones
(Statins). Example of inhibiting hormones: Dopamine (inhibiting hormone for prolactin),
somatostatin (for GH). These neurons (which secrete releasing & inhibitory hormones)
originate in various parts of the hypothalamus and send their nerve fibers to the median
eminence and tuber cinereum, an extension of hypothalamic tissue into the pituitary stalk.
Their function is not to transmit signals from one neuron to another, but rather to secrete the
hypothalamic releasing and inhibitory hormones. These hormones are immediately absorbed
into the hypothalamic-hypophysial portal system and carried directly to the sinuses of the
anterior pituitary gland.
Adenohypophysis is made up of cords of cells; among them there are fenestrated
capillaries. The secretion of these cells is released directly into the blood. According to the

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staining property, the cells of anterior pituitary are chromophobe (50% of them, they are not
secretory in nature) and chromophil cells (they contain large number of granules and are
darkly stained ; there are two types - acidophilic and basophilic cells).
From morphological point of view, there have been identified several types of cells:
 Corticotropes cells (basophils) - synthesize Adrenocorticrotrope hormone
(ACTH)
 Thyrotropes, synthesize thyroid stimulating hormone (TSH) (basophils)
 Somatotropes: Growth hormone (GH) (acidophils)
 Lactotropes: Prolactin (PRL) (acidophils)
 Gonadotropes: gonadotrophins - FSH and LH (basophils)
TSH is a glycoprotein (31000 D). This hormone stimulates the secretion of thyroid
hormones: Tyroxin (T4) and triioiodotyronin (T3); it also stimulates the development (increase
in size) of the thyroid gland.
FSH (30000 D) is a glycoprotein made up of two subunits, alpha and beta, produced by
gonadotrope cells. Its life time is about 3-4 hours. This hormone is synthesized beginning with
the puberty, and there is a cyclical secretion of it. It stimulates the development of follicle de
Graaf in women and secretion of estrogen by the theca cells of graafian follicle. In man this
hormone stimulates spermatogenesis.

Secretion of pituitary hormones (after A. Despopoulos, S. Silbernagl, Color Atlas of Physiology,

2003)

LH also a gonadotrope hormone; its life time is an hour. In women it stimulates

ovulation, causes maturation of vesicular follicle into graafian follicle along with FSH. It is

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necessary for the formation of corpus luteum. In males it stimulates Leydig cells to produce
testosterone.
TSH, FSH and LH are glycoproteins. They are made up of 2 subunits: alpha and beta.
Alpha subunit is common for all of them, while beta one is specific to each of them. Only when
these two subunits are together, these hormones are active.
ACTH is made up of 39 Aa, having the molecular weight around 4500 D. It is
synthesized by corticotrope cells. It stimulates the secretion of glucocorticoid hormones
(cortisol) at the level of adrenal glands. Secretion of ACTH is highest in the morning and lowest
in the evening, so there is a circadian variation in the secretion of this hormone. ACTH is
synthesized as a pro-hormone, named proopio-melanocortin (POMC). POMC is made up of
several fragments:
1) N-terminal fragment
2) ACTH
3) Beta lipotropin
Beta lipotropin stimulates the secretion of aldosterone. By its cleavage another two
molecules are released gamma lipotropin and beta endorphin. Beta lipotropin in animals
produces lipolysis and mobilizes fatty acids from tissues. Breaking down of N-terminal
fragment produces alpha melanocyte stimulating hormone (MSH). Because ACTH contains
the alpha MSH amino acid sequence at its N-terminal end, it has melanocyte-stimulating
activity when present in blood at high concentrations. Humans who have high blood levels of
ACTH, as a result of Addison’s disease or an ACTH-secreting tumor, are often hyperpigmented.
In the hypothalamus, alpha MSH is important in the regulation of feeding behavior.
Prolactin is a polypeptide having the molecular weight of 26000 D (199 aa). It has a
similar structure to growth hormone. It is synthesized by lactotropes. It stimulates the
development of mammary gland and milk secretion. It causes localized alveolar hyperplasia
of mamary gland during pregnancy. Prolactin secretion starts increasing with the 5 th month of
pregnancy. The synthesis and secretion of PRL is stimulated by estrogens and other hormones,
such as TRH, which increases the expression of the PRL gene. Dopamine inhibits the synthesis
of PRL.
GH is a polypeptide with the molecular weight of 21,500 D and 191 aa; It is produced
by somatotropes; it is similar in structure with prolactin. It is transported in blood by GH
binding proteins (GHBPs). GH stimulates the growth of the body. This action is due to the
stimulation of protein synthesis, production of nucleic acids, cell division. GH has two
important effects :
1) stimulation of the growth of the organism
2) metabolic effects

 Stimulation of growth by GH

It causes growth of almost all tissues of the body that are capable of growing. It
promotes increased sizes of the cells and increased mitosis, with development of greater
numbers of cells and specific differentiation of certain types of cells such as bone growth cells
and early muscle cells. The most important effect is increase in length of bones. This results
from : (1) increased deposition of protein by the chondrocytic and osteogenic cells, (2)
increased rate of reproduction of these cells, and (3) a specific effect of converting
chondrocytes into osteogenic cells, thus causing deposition of new bone.
There are two principal mechanisms of bone growth:

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 1) in response to growth hormone stimulation, the long bones grow in length at the
epiphyseal cartilages, where the epiphyses at the ends of the bone are separated from the
shaft. This is also called epiphyseal growth plate. This growth first causes deposition of new
cartilage, followed by its conversion into new bone, thus elongating the shaft and pushing the
epiphyses farther and farther apart. Osteoblasts, the bone-forming cells, at the shaft edge of
the epiphyseal growth plate convert the cartilaginous tissue at this edge to bone while new
cartilage is simultaneously being laid down in the interior of the plate by cells called
chondrocytes. At the same time, the epiphyseal cartilage itself is progressively used up, so
that by late adolescence, no additional epiphyseal cartilage remains to provide for further long
bone growth. At this time, bony fusion occurs between the shaft and the epiphysis at each
end, so that no further lengthening of the long bone can occur. This is known as epiphyseal
closure and occurs at different times in different bones.
Thus, growth hormone promotes bone lengthening by stimulating maturation and cell
division of the chondrocytes in the epiphyseal plates, thereby continuously widening the
plates and providing more cartilaginous material for bone formation.
2) osteoblasts in the bone periosteum and in some bone cavities deposit new bone on
the surfaces of older bone. Simultaneously, osteoclasts in the bone remove old bone. When
the rate of deposition is greater than that of resorption, the thickness of the bone increases.
Growth hormone strongly stimulates osteoblasts. Therefore, the bones can continue to
become thicker throughout life under the influence of growth hormone; this is especially true
for the membranous bones. For instance, the jaw bones can be stimulated to grow even after
adolescence, causing forward protrusion of the chin and lower teeth. Likewise, the bones of
the skull can grow in thickness and give rise to bony protrusions over the eyes.
At the level of different cells of the body there are receptors for growth hormone, so
when it reacts with its receptor, some substances are synthesized which are similar in
structure with insulin. This is why they are named insulin-like growth factors. It is accepted
today that liver cells, chodronblasts, osteoblasts, fibroblasts have receptors for growth
hormone, and GH stimulates the synthesis of insulin-like growth factors I and II (IGFs). These
substances (IGFs) stimulate the synthesis of collagen and introduce sulfate groups into the
structure of cartilages.
Initially IGFs were called somatomedins (A,B,C,D). But later it has been identified that
somatomedin C is the same as Insulin-like growth factor I (IGF). IGF I is similar in structure
with proinsulin. The molecular weight of somatomedin C is about 7500 D, and its
concentration in the plasma closely follows the rate of growth hormone secretion. GH - life
time = 20 minutes, somatomedin C - is bound by a carrier protein in plasma and slowly
released to tissues - life time = 20 hours.
Because insulin-like growth factors have some insulinic effects (produce
hypoglycemia), insulin mediates its role in growth because of these insulin-like growth factor.
Besides GH, the synthesis of Insulin-like growth factors is changed by other substances; for
instance glucocorticoid hormones and hypoproteinemia and increase concentration of
oestrogens inhibit the production of insulin-like growth factor I. IGF stimulates the division of
chondrocytes, the synthesis of RNA and DNA, the formation of collagen, favorising the
convertion of prolin into hydroxyl-prolin, and by this they stimulate the development of
growth plate and the increase in length of bone.
The production of Gh is inhibited by the negative feedback of IGF. At the level of
hypothalamus, it also inhibits the release of GH releasing factor.

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  Metabolic effects of GH:

GH increases the permeamibility of cell membrane for amino acids (aa), increasing the
transport of amino acids through the cell membrane. It stimulates the synthesis of proteins
and nuclei acids (RNA, DNA, Ribosomal nuclei acid). GH increases RNA translation, causing
protein to be synthesized in greater amounts by the ribosomes, increases nuclear
transcription of DNA to form RNA. It decreases the catabolism of proteins and Aa (it is a
protein sparer- fats are used as source of energy).
It stimulates lypolysis, and by this fatty acids are mobilized into the circulation,
followed by the oxidation of fatty acids. Fatty acids are used as source of energy by the cells.
GH is a ketogenic hormone, involved in the production of ketone bodies. Sometimes excess
mobilization of fats from the adipous tissue causes accumulation of fats in liver, resulting in
fatty liver.
GH has a diabetogenic effect, it produces hyperglycemia. This is done by decrease of
using of glucose by cells for the production of energy. Acetyl CoA formed during the
metabolism of fat inhibits the glycolytic pathway. GH increases the deposition of glycogen in
the cells. Because of this no glucose enters the cells. Ultimately glycemia increases. Glucose is
released from liver. It stimulates insulin secretion directly and because increases glycemia.
Due to the excess stimulation of insulin secretion, the beta cells are burn out, which leads to
diabetes mellitus. Somatotropic hormone decreases the number of receptors for insulin, and
inhibits the phosphorylation of glucose inside the cells (= first step of glycolysis).
GH increases the intestinal absorption of Ca2+ and phosphate and decreases renal
excretion of sodium and potassium.

Factors stimulating the secretion of growth hormone: The secretion of Gh is not the
same troughout the day. There are several peaks, 3 to 4 times a day, lasting about 20-30
minutes, when growth hormone secretion is higher. The highest secretion is noted during the
sleep, specially after midnight (between midnight and 4 o’clock in the morning). The lowest
value is reached in the afternoon, around 4 o’clock. The blood concentration of Gh in adult is
between 2-3 ng/mL, in a child or adolescent is 5-6 ng/mL. After adolescence, secretion
decreases slowly with aging, finally falling to about 25 per cent of the adolescent level in very
old age.
The secretion of growth hormone is stimulated by several factors: starvation,
hypoglycemia (below 70 mg/dL); increased concentration of amino acids: Arginine, Leucine,
Tryptophane, another factors are: stress, trauma, physical effort, sleep, especially the 3rd and
the 4th phase of sleep. The secretion of GH is decreased in obese persons, hyperglycemia,
increase of FFA in blood.

Physiology of growth

Growth of the organism is accelerated in the first 2 years of life and at puberty. The
growth is determined by genetical, external (environmental factors) and endocrine factors.
a) Genetical factors: children of two short parents will be probably short.
b) External factors: nutrition - it is recommended that animal proteins (especially in
childhood) to be found in diet of children; diseases during childhood - it has been noted that
secretion of cortisol is increased in these periods. Cortisol stimulates the catabolism of
protein. By this the growing is delayed. But after the disease is cured, the growing is

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accelerated until the growth reaches the normal value. It is considered that emotional
deprivation and psychological factors can impair growth by complex, poorly understood
mechanisms.
c) Endocrine factors: several hormones stimulate the growth. Thyroid hormones (T3,
T4) stimulate the calcification of cartilages, development of teeth, and the development of a
normal aspect of face. Besides this, T3 and T4 stimulate the secretion of GH, because they are
required for both the synthesis of growth hormone and the growth-promoting effects of that
hormone. When there is a deficiency of thyroid hormones the organism is not developed
properly, the person is very short, the body is not proportional and this is related with a mental
retardation. By comparison to pituitary dwarfism (due to the lack of GH - the person looks
normal, and there is no mental retardation), in case of thyroid dwarfism - the person is short,
very disarmonic and mental retarded.
Insulin exerts direct, specific growth-promoting effects on cell differentiation and
cell division during fetal life (and possibly during childhood). Moreover, insulin is required for
normal production of IGF-I. In children suffering from diabetes mellitus, if they do not get
insulin, they remain shorter.
Sexual hormones (testosteron) stimulate the secretion of IGF I. The level of this
hormone is very high during childhood, and the maximum is reached at the age of 13-17 years.
Testosteron initially stimulates the growing of bones, but after a while it determines the
ossification of growth cartilages, and the growth stops.
Oestrogens in reduced concentration stimulate growth, while in high concentrations,
inhibit the growth. Oestrogen favorizes the secretion of GH. Increased concentration of
oestrogen inhibits secretion of insulin-like growth hormone. Glucocorticoid hormones have a
negative impact on growth. Cortisol, when present in high concentration, inhibits DNA
synthesis and stimulates protein catabolism in many organs. Moreover, it causes bone
breakdown by inhibiting osteoblasts and stimulating osteoclasts. It also inhibits the secretion
of growth hormone. For all these reasons, in children, the elevation in plasma cortisol that
accompanies infections and other stresses is, at least in part, responsible for the retarded
growth that occurs with illness.
The cells of many tissues and organs of the body have receptors for GH in their plasma
membranes. The interaction of GH with these receptors produces its growth-promoting and
other metabolic effects, but the mechanisms that produce these effects are not fully
understood. The binding of GH to its receptor activates a tyrosine kinase (JAK2), which initiates
changes in the phosphorylation pattern of cytoplasmic and nuclear proteins. These
phosphorylated proteins ultimately stimulate the transcription of specific genes, such as that
for IGF-I.
Many of the mitogenic effects of GH are mediated by IGF-I; however, evidence
indicates that GH has direct growth-promoting actions on progenitor cells or stem cells, such
as prechondrocytes in the growth plates of bone and satellite cells of skeletal muscle. GH
stimulates such progenitor cells to differentiate into cells with the capacity to undergo cell
division. An important action of GH on the differentiation of progenitor cells is stimulation of
the expression of the IGF-I gene; IGF-I is produced and released by these cells. IGF-I exerts an
autocrine mitogenic action on the cells that produced it or a paracrine action on neighboring
cells. In response to IGF-I, these cells undergo division, causing the tissue to grow mainly
through cell replication.

Regulation of adenohypophyis secretion

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 It is under the control of hypothalamus by means of releasing hormones and inhibiting
hormones. There are known several releasing hormones:
 Corticotropin releasing hormone (CRH) for adrenocorticotrope
hormone/adrenocorticotropin (ACTH); it has 41 aa and it is secreted by the neurons
from paraventricular nuclei of hypothalamus (not the same producing oxytocin and
ADH); these neurons are small and are named parvicellular cells. The receptor for CRH
is coupled with G protein and it activates adenylyl cyclase, which catalyzes the
formation of cAMP from ATP.
 Thyrotropin releasing hormone (TRH): it has 3 aa, it stimulates the secretion of thyroid
stimulating hormone (TSH).
 GH releasing hormone (GHRH): made up of 44 aa, stimulates the secretion of GH. It is
synthesized by ventromedial and arcuate nuclei of Ht. By coupling with its receptor on
somatotropes the conc. of cAMP increases, that leads to increase of Ca 2+ and
exocytosis of vesicles with hormones, but also synthesis of new molecules of hormone.
Somatostatin (growth hormone inhibitory hormone (GHIH – 14 aa) inhibits release of
GH. It has been also identified in gastric mucosa, intestinal mucosa, pancreas, posterior
horns of spinal cord and hypophysis. It inhibits the secretion of Gh, TSH, insulin,
glucagon, gastrin, secretin, colecystokinine, vasoactive intestinal peptide. It increases
gastric secretion and pancreatic secretion, it decreases the motility of stomach and
biliary tract. Therapeutically it is used for treatment of persons having pituitary tumors
that secrete growth hormone. Expression of GH gene is also stimulated by thyroid
hormones. The thyroid hormones increase the release of somatostatin from
Hypothalamus.
 Gonadotropin releasing hormone (GRH) - 10 aa : stimulates the secretion of FSH and
LH.
 Prolactin releasing hormone (PRH)- stimulates prolactin secretionm. It exists also PIRH,
a prolactin inhibiting releasing hormone. It is has been identified as being dopamine.

Neurohypophysis

The posterior pituitary gland, also called the neurohypophysis, is composed mainly of
glial-like cells called pituicytes. The pituicytes do not secrete hormones; they act as a
supporting structure for large numbers of terminal nerve fibers from nerve tracts that
originate in the supraoptic and paraventricular nuclei of the hypothalamus. It is not a gland,
because it does not secrete hormones, but hormones are stored and released at this level.
These hormones are synthesized by paraventricular and supraoptic nuclei of hypothalamus.
These neurons are magnocellular cells. At their level ADH and oxytocin are secreted together
with neurophysin type I and type II, that are parts of precursor molecule - neurophysin II with
ADH and oxytocin with neurophysin I.
ADH is formed primarily in the supraoptic nuclei, whereas oxytocin is formed primarily
in the paraventricular nuclei. Each of these nuclei can synthesize about one sixth as much of
the second hormone as of its primary hormone. During the axoplasmic transport, ADH is
separated from neurophysin II and oxytocin from neurophysin I. At the level of terminal knobs,
there will be found vesicles with free ADH, free oxytocin, free neurophysin I and free
neurophysin II.

12
  ADH (antidiuretic hormone/vasopressin)

It is a polypeptide (9 aa)with a life time of 18 –20 minutes. In human beings ADH is also
called vasopressin or arginine vasopressin, while in other species lysine vasopressin is found.
ADH acts on 2 types of receptors: RV1 and RV2. RV1 is found at level of smooth muscle cells
of blood vessels. The action of ADH on this receptors is mediated by a G-protein (IP3 and DAG)
and vasoconstriction is the result. RV2 is found in distal convuluted tubule and collecting ducts
of nephrons. This action is mediated by adenylyl cyclase. Vesicles with aquaporines 2 will be
inserted on cell membrane of tubular cells and water is reabsorbed. The main role of ADH is
to stimulate reabsorption of water in collecting tubule of kidney (to preserve water in the
organism = antidiuresis).

Regulation of ADH secretion

When the osmolarity of blood is above 285 mOsm/L the secretion of ADH is
stimulated, together with the thirst sensation. The precise way that the osmotic concentration
of the extracellular fluid controls ADH secretion is not clear. Somewhere in or near the
hypothalamus are modified neuron receptors called osmoreceptors. When the extracellular
fluid becomes too concentrated, fluid is pulled by osmosis out of the osmoreceptor cells,
decreasing their size and initiating appropriate nerve signals in the hypothalamus to cause
additional ADH secretion. Conversely, when the extracellular fluid becomes too dilute, water
moves by osmosis in the opposite direction, into the cell, and this decreases the signal for ADH
secretion. Although some researchers place these osmoreceptors in the hypothalamus itself
(possibly even in the supraoptic nuclei), others believe that they are located in the organum
vasculosum, in the anteroventral wall of the third ventricle.
The distention of atria and great vessels (stretch receptors/voloreceptors) by
increasing of blood volume will inhibit the secretion of ADH. When the blood volume
increases, there is more blood coming to atria. By this the wall of atria will be distended and
by a reflex mechanism the secretion of ADH is inhibited. The distension of atria also releases
Atrial Natriuretic Peptide (ANP), which inhibits the release of ADH.
Baroreceptors influence the secretion of ADH. When blood pressure decreases, by a
reflex mechanism, ADH secretion is stimulated.
Exposure of the organism to cold - in this case, there is a peripheral vasoconstriction
to prevent heat loss; this is associated with mobilization of blood towards internal organs,
increase of venous return, and by this atria are distended, and the same reflex mechanism
inhibits the secretion of ADH.
Angiotensin 2 : stimulates the secretion of ADH .
Stress, emotions, some drugs (morphine, nicotine, barbiturates) increase the
secretion of ADH. Alcohol and other drugs inhibit the secretion of ADH.
Decrease of blood volume stimulates the secretion of ADH.
 Oxytocin

It has a molecular weight of 1 000 D, it is made up of 9 aa and it has a life time of 6

mintes. Oxytocin performes several roles:
o On mammry glands in women - It causes the ejection of milk, because it
produces the contraction of myoepithelial cells from the ducts of the mammary gland. This

13
mechanism works as follows: the suckling stimulus the touch receptors around the nipple and
causes signals that are transmitted through sensory nerves to the oxytocin neurons in the
paraventricular and supraoptic nuclei in the hypothalamus. This induces the release of
oxytocin by the posterior pituitary gland. The oxytocin is then carried by the blood to the
breasts, where it causes contraction of myoepithelial cells that lie outside of and form a
latticework surrounding the alveoli of the mammary glands. In less than a minute after the
beginning of suckling, milk begins to flow. This mechanism is called milk letdown or milk
ejection. It is a positive feedback mechanism.
o On pregnant uterus – the hormone produces the contraction of pregnant
uterus during delivery. At the onset of labor when cervix is dilated, the fetus descends through
the birth canal. The receptors from the cervix are stimulated and impulses are transmitted to
the hypothalamus, followed by the stimulation of oxytocin secretion. Oxytocin is released by
means of positive feedback mechanism. It induces contraction of uterus, which in turn causes
release of higher amounts of oxytocin. During labour the concentration of oxytocin increases
several times, from 50 microunits/mL until 150 microunits/mL. In animals having removed
pituitary gland the labour is prolonged. Throughout the period of pregnancy oxytocin
secretion is inhibited by estrogen and progesteron. At the end of pregnancy the secretion of
these two hormones decreases and the secretion of oxytocin is not more inhibited. During the
later stages of pregnancy the number of R for Oxytocin in the wall of uterus increases. Due to
this the uterus becomes more sensitive to hormone. The hormone also stimulates the
secretion of Prostaglandins by placenta, that will intensify the uterine contractions induced by
oxytocin.
o On nonpregnant uterus – oxytocin is also involved in fertilization (meeting of
ovum with spermatozoa). During intercourse, the recepetors from vagina are stimulated. This
generates impulses that are transmitted to the hypothalamus, increasing the secretion of
oxytocin. Oxytocin is released into circulation and reaches the female genital tract. Here are
produced antiperistaltic contractions of uterus that help spermatozoa to progress towards
oviduct .
o In males – the secretion of oxytocin increases during ejaculation. This causes
contraction of smooth muscle fibers especially in vas deferens and facilitates the release of
sperm into urethra.

Disturbances of pituitary gland secretion

Panhypopituitarism – can be congenital or inherited (tumors of pituitary gland,

surgery, infarction of hypophysis). This is followed by atrophy of adrenal gland, thyroid gland
and gonads, due to decreased secretion of all hormones of adenohypophysis.
Diabetes insipidus - it is the disease produced by decreased secretion of ADH. This is
the result of destruction of supraoptic and paraventricular nuclei (tumors, cerebral
traumatisms, infection, stroke) or the kidney does not responde to ADH (nephrogenic diabetes
insipidus). Water is not reabsorbed, so until 12 - 20 liters of urine can be lost daily.
SIADH ( Syndrom of Inappropriate Hypersecretion of ADH) is a disease characterized
by loss of Na+ through urine due to the hypersecretion of ADH (cerebral tumors, lung tumors
- the tumoral cells secrete ADH or ADH- like peptide). ADH normaly stimulates the reabs.of
water in collecting tubule and the urine that is excreted is concentrated with sodium and other
ions. Loss of Na+ decreases plasma osmolarity. The hypotonic plasma inhibits the secretion of

14
ADH. In SIADH the secretion of ADH from tumor cells is not inhibited by hypotonic plasma, so
there is a continuos loss of Na+, resulting in persistent hypotonicity of plasma.
Another pathological condition is the lack of secretion of GH during childhood, which
leads to pituitary dwarfism. Treatment - with human GH, obtained by recombinant DNA
technology. A person with panhypopituitary dwarfism does not pass through puberty and
never secretes sufficient quantities of gonadotropic hormones to develop adult sexual
functions. In one third of such dwarfs, however, only growth hormone is deficient; these
persons do mature sexually and occasionally reproduce.
An excessive secretion of GH during childhood will induce a disease named gigantism.
It is due to a tumor secreting GH. Hyperglycemia and DM may be associated and finally
panhypopituitarism occurs (if they are not treated) - because the tumor destroys the
hypophysis and all hormones are not secreting anymore. Acromegaly - hypersecretion of GH
during adulthood due to the adenomatous tumors of acidophil cells from anterior pituitary.
Many times diabetes mellitus is associated, together with some neurological, ophtalmological
signs, kyphosis. Many soft tissues organs (the tongue, the liver, and especially the kidneys),
become greatly enlarged. Enlargement is especially marked in the bones of the hands and feet
and in the membranous bones, including the cranium, nose, bosses on the forehead,
supraorbital ridges, lower jawbone, and portions of the vertebrae, because their growth does
not cease at adolescence.

THYROID GLAND

Thyroid gland is bigger than pituitary, it has 25 – 30 g. It is made up of two lobes,

connected through the isthmus, which lies just below the cricoid cartilage. Thyroid is larger in
females than in males. Its function increases during pregnancy and lactation and decreases
during menopause. Thyroid galnd strats functioning in the fetal life, but the maximum activity
of the gland is reached only after puberty.
The structural and functional unit of thyroid gland is thyroid follicle. There are around
3 million of follicles, having the size between 100-500 micrometers. These follicles are made
up of a layer of cuboid cells, surrounded by a basement membrane. The apical membrane of
follicle cells is covered with microvilli. The lateral membranes of the follicular cells are
connected by tight junctions, which provide a seal for the contents of the lumen. The basal
membrane of the follicular cells is close to the rich capillary network that penetrates the
stroma between the follicles. According to the state of activity of thyroid gland, the follicles
may be smaller and the cells become columnar (when the gland is active) or, the follicles are
bigger and the cells are flattened, with much colloid (when the gland is inactive). Inside of
follicles there is a thick, gel-like substance named colloid. The colloid is a solution composed
primarily of thyroglobulin, a large protein that is a storage form of the thyroid hormones.
Outside of the cells of follicles, but above the basement mb., there is one more type of
cells named parafolliclular cells or C-cells, which synthesize calcitonin, involved in calcium
metabolism. The thyroid hormones are thyroxin (T4) and triiodothyronin (T3).

Synthesis of the thyroid hormones

15
 This process involves the synthesis of a thyroglobulin precursor, the uptake of iodide,
and the formation of iodothyronine residues. The cells of follicle secrete two hormones - T3
and T4. Both T3 and T4 are organic compounds containing iodine.
Iodine is found in food and is absorbed as iodide. Daily intake of Iodine is 100 - 500
micrograms, which is in balance with the amount of iodine which is lost through urine (100 –
150 μg) and feces (20 μg). Thyroid gland takes up daily around 120 μg of iodine and releases
in circulation around 80 micrograms of iodine under the form of thyroxin and triiodothyroin.
Iodine is an essential element that functions as a component of T4 and T3. Most of the iodine
ingested in food is absorbed into the blood from the gastrointestinal tract by active transport;
in the process it is converted to the ionized form, iodide. Iodide is actively transported from
the blood into the thyroid follicular cells. Once in the cells, the iodide is converted back to
iodine, which is then coupled to the side chains of tyrosine molecules that had previously been
incorporated into thyroglobulin precursor. Iodine is also concentrated by salivary glands,
stomach, mammary gland , placenta, eye ball.
The iodine used for iodination of the thyroglobulin comes from the blood perfusing the
thyroid gland. The basal plasma membranes of follicular cells, which are near the capillaries
that supply the follicle, contain iodide transporters (a sodium –iodide symport pump). These
transporters move iodide along with sodium across the basal membrane and into the cytosol
of the follicular cell. The iodide transporter is an active transport mechanism that requires
ATP, is saturable, and can also transport certain other anions, such as bromide, thiocyanate,
and perchlorate. This mechanism enables the follicular cell to concentrate iodide many times
over the concentration of iodide present in the blood (30X); therefore, follicular cells are
efficient extractors of the small amount of iodide circulating in the blood. Once inside follicular
cells, the iodide ions diffuse rapidly to the apical membrane, where they are used for
iodination of the thyroglobulin precursor.
The first step in the formation of the thyroid hormones is conversion of the iodide ions
to an oxidized form of iodine, that then combines directly with the amino acid tyrosine. This
oxidation of iodide is catalyzed by the enzyme peroxidase. The peroxidase is either located in
the apical membrane of the cell or attached to it.
The synthesis of the protein precursor for thyroglobulin is another step in the formation
of T4 and T3. This substance is a 660 kDa glycoprotein composed of two similar 330 kDa
subunits held together by disulfide bridges. The subunits are synthesized by ribosomes on the
rough ER and then undergo dimerization and glycosylation in the smooth ER. The completed
glycoprotein is packaged into vesicles by the Golgi apparatus. These vesicles migrate to the
apical membrane of the follicular cell and fuse with it. The thyroglobulin precursor protein is
then extruded onto the apical surface of the cell, where iodination takes place.
The next step in the formation of thyroid hormones is the addition of one or two iodine
atoms to certain tyrosine residues of thyroglobulin. The precursor of thyroglobulin contains
134 (140) tyrosine residues, but only a small fraction of these become iodinated. A
thyroglobulin molecule contains only 20 to 30 atoms of iodine. The iodination of thyroglobulin
is catalyzed by the enzyme thyroid peroxidase, which is bound to the apical membranes of
follicular cells. Thyroid peroxidase binds an iodide ion and a tyrosine residue in the
thyroglobulin precursor, bringing them in close proximity. The enzyme oxidizes the iodide ion
and the tyrosine residue to short lived free radicals, using hydrogen peroxide that has been
generated within the mitochondria of follicular cells. The free radicals then undergo addition.
The product formed is a monoiodotyrosine (MIT) residue, which remains in peptide linkage in
the thyroglobulin structure. A second iodine atom may be added to a MIT residue by this same

16
enzymatic process, forming a diiodotyrosine (DIT) residue. The binding of iodine with the
thyroglobulin molecule is called organification of the thyroglobulin.

Structure of T4 and T3 ( after A.C. Guyton & J. Hall, Textbook of Medical Physiology,
2006)

Iodinated tyrosine residues that are close together in the thyroglobulin precursor
molecule undergo a coupling reaction, which forms the iodothyronine structure. Thyroid
peroxidase is believed to catalyze the coupling reaction through the oxidation of neighboring
iodinated tyrosine residues to short-lived free radicals. These free radicals undergo addition.
The addition reaction produces an iodothyronine residue and a dehydroalanine residue, both
of which remain in peptide linkage in the thyroglobulin structure. One molecule of MIT
combines with one molecule of DIT to form tri-iodothyronine (T3). When two neighboring DIT
residues couple, T4( tetra-iodothyronine or thyroxine) is formed. Sometimes one molecule
of MIT and one molecule of DIT combine to produce another form of T3 named reverse T3 or
rT3. This is biological inactive.
Only about 20 to 25% of the DIT and MIT residues in the thyroglobulin molecule become
coupled to form iodothyronines. A thyroglobulin molecule contains five to six uncoupled
residues of DIT and two to three residues of T4. T3 is formed in only about one of three
thyroglobulin molecules. The thyroid gland secretes more T4 than T3.
When the thyroid gland is stimulated to secrete thyroid hormones, pinocytosis occurs
at the apical membranes of follicular cells. Pseudopods from the apical membrane reach into
the lumen of the follicle, engulfing bits of the colloid. Endocytotic vesicles or colloid droplets
formed by this pinocytotic activity migrate towards the basal region of the follicular cell.
Lysosomes fuse with the colloid droplets and hydrolyze the thyroglobulin to its constituent
amino acids. As a result, T4 and T3 and the other iodinated amino acids are released into the
cytosol.

17
 T4 and T3 formed from the hydrolysis of thyroglobulin are released from the follicular
cell and enter the capillary circulation. The DIT and MIT generated by the hydrolysis of
thyroglobulin are deiodinated in the follicular cell by iodotyrosine deiodinase. They are not
released into the circulation. The released iodide is then reutilized by the follicular cell for the
iodination of thyroglobulin.

Thyroid hormone synthesis and secretion (Rhoades R, Tanner G, Medical Physiology, second
edition, Lippincott Williams & Wilkins, 2003)

About 93% of the thyroid hormone released from the thyroid gland is normally
thyroxine (T4) and only 7% is triiodothyronine (T3). After entering the peripheral tissues
(liver, kidney, muscles) most of T4 is deiodinated to T3. It is believed that the true intracellular
hormone is T3 rather than T4.

Transport and metabolism of Th

Most of the T4 and T3 molecules that enter the blood tream become bound to plasma
proteins. About 70% of the T4 and 80% of the T3 are noncovalently bound to thyroxine-
binding globulin (TBG), a 54-kDa glycoprotein that is synthesized and secreted by the liver.
Each molecule of TBG has a single binding site for a thyroid hormone molecule. The remaining
T4 and T3 in the blood are bound to transthyretin (15 –20%) or to albumin (9%). Less than 1%
of the T4 and T3 in blood is in the free form, and it is in equilibrium with the large protein-
bound fraction. 99.96% of T4 is bound and 0.04% is free; 99.6% of T3 is bound and 0.4% is
free.
The protein-bound form of T4 and T3 represents a large reservoir of preformed
hormone that can replenish the small amount of circulating free hormone as it is cleared from
the blood. This reservoir provides the body with a buffer against changes in circulating thyroid
hormone levels as a result of sudden changes in the rate of T4 and T3 secretion. The protein-

18
bound T4 and T3 molecules are also protected from metabolic inactivation and excretion in
the urine. As a result of these factors, the thyroid hormones have long half-lives in the
bloodstream. The half-life of T4 is about 7 days; the half-life of T3 is about 1 day (10 –24
hours).

Metabolism of Th- by : - deiodination ( T4 is transformed into T3, rT3, T2, T1, T0- these
compounds do not have biological activity)
- conjugation with sulphate and glucuronate and secretion in bile
- decarboxilation and deamination to TRIAC ( triiodo thyroactic
acid) and TETRAC (tetraiodo –thyroacetic acid)
- decarboxilation to T3 amine and T4 amine

Effects/ functional roles of Th ( thyroid hormones)

Th act by activating the genes and stimulating the synthesis of proteins and RNA at
mitochondrial level. The receptor for Th is either attached to DNA or in closed proximity to it.
Th perform several functions/ effects:
- they increase the oxygen consumption and the metabolic activity of cells (muscles,
heart, liver , kidney – these organs have many Th receptors) ; other tissues - brain, lymphoid
organs, skin, spleen, retina, gonads - have a little response to Th. BMR increases up to 60 –
100% above normal under the action of high amounts of Th (thyroid hormones). BW ( body
weight) decreses, but this can be counterbalanced by increasing the apetite under the action
of Th. In hypothyroidism BW rises up.
- Th increase the number, size and activity of mitocondria in cells. These produce more
ATP.
- they increase the activity of Na/K- ATP-ase , having as result production of more heat
= thermogenic action/calorigenic effect. Thyroid hormones stimulate the synthesis of
uncoupling protein-1 (UCP-1) in brown adipose tissue. ATP is synthesized by ATP synthase in
the mitochondria when protons flow down their electrochemical gradient. UCP-1 acts as a
channel in the mitochondrial membrane to dissipate the ion gradient without making ATP. As
the protons move down their electrochemical gradient uncoupled from ATP synthesis, energy
is released as heat. Adult humans have little brown adipose tissue, so it is not likely that UCP-
1 makes a significant contribution to nutrient oxidation or body heat production. However,
several uncoupling proteins (UCP-2 and UCP-3) have recently been discovered in many tissues,
and their expression is regulated by thyroid hormones.
- Th promote growth and development of the brain during fetal life and for the first few
years of postnatal life. If the fetus does not secrete sufficient quantities of both before birth
and afterward are greatly retarded, and the brain remains smaller than normal. Without
specific thyroid therapy within days or weeks after birth, the child without a thyroid gland will
remain mentally deficient throughout life. Animal studies have demonstrated that thyroid
hormones inhibit nerve cell replication in the brain and stimulate the growth of nerve cell
bodies, the branching of dendrites, and the rate of myelinization of axons. These effects of
thyroid hormones are presumably due to their ability to regulate the expression of genes
involved in nerve cell replication and differentiation. Thyroxine also stimulates the blood flow
to brain and is a stimulating factor for the CNS (brain).

19
 - T4 is essential for the normal activity of skeletal muscles. Slight increase of T4 makes
muscles to contract more vigorous. In hyperthyroidism  weakness of the muscles (due to
the catabolism of proteins), fine muscular tremor (10 – 15 times/sec). The lack of Th makes
muscles more sluggish.
- Th promote the body growth by stimulating the expression of GH gene in the
somatotrophs in the anterior pituitary gland; therefore, a thyroid hormone-deficient
individual will also be GH-deficient. If this condition occurs in a child, it will cause growth
retardation. Thyroid hormones also promote the calcification and, hence, the closure, of the
cartilaginous growth plates of the bones of the skeleton. This action limits further linear body
growth.
- Th stimulate the synthesis and breaking down of proteins, the effect depends on
dose. Decreased level of Th are protein anabolic, while increased doses are catabolic and
promoting of urinary excretion of nitrogen. Protein turnover is favorized by Th. Th stimulate
the penetration of aa in cells and their incorporation in new proteins. Th increase the synthesis
of proteins in cells by stimulating the translation of RNA, increasing the transcription of DNA
to RNA, by increasing the activity of mitochondria and the activity of cellular enzymes.
- Th stimulate carbohydrate metabolism: increase uptake of glucose by the cells,
enhance glycogenolysis, enhance gluconeogenesis, increase rate of absorption from the
gastrointestinal tract, increase insulin secretion.
- lipids are mobilized from the fat tissue, with the increase of plasma conc. of free fatty
acids and their oxidation by cells. Th decrease the conc. of cholesterol (secretion of Ch in bile
is stimulated by Th; Th induces increased number of LDL- receptors on the liver cells, leading
to rapid removal of LDL from plasma and secretion of Ch into bile), phospholipids and Tg in
plasma. They stimulate deposition of fats in the liver  fatty liver.
- Th increase the need for vitamins, because they stimulate enzyme activity.
- Th rise CO and blood flow, the strength of heart contraction, heart rate , systolic BP,
but decrease diastolic BP. Thyroid hormones enhance the action of adrenaline and
noradrenaline.
- the gastrointestinal motility is increased together with digestive secretions (diarrhoea
in hyperthyroidism, constipation in hypothyroidism). Also appetite and food intake is enhaced.
- Th increase the rate and force of respiration (due to the increased demand of O2 and
production of CO2).
- they have excitatory effects on nervous and muscular activity (muscle tremor),
difficulty to sleep.
- they influence sexual function - lack of Th  loss of libido, menorrhagia,
polymenorrhea or amenorrhea.
- they stimulate the disssociation of oxyhemoglobin, due to the increase of 2,3 DPG in
RBCs.
- Th induce conversion of caroten in vit A in liver
- Th stimulate erythropoiesis. In hyperthyroidism  polycythemia.

Regulation of thyroid gland secretion

The thyroid hormones regulate their own secretion. T3 exerts an inhibitory effect on
TSH secretion by thyrotrophs in the anterior pituitary gland by decreasing thyrotrophs
sensitivity to thyrotropin-releasing hormone (TRH). The activity of TRH on pituitary cells is

20
mediated by phospholipase C. When the circulating concentration of free thyroid hormones
is high, thyrotrophs are relatively insensitive to TRH, and the rate of TSH secretion decreases.
The resulting fall of TSH levels in the blood reduces the rate of thyroid hormone release from
the follicular cells in the thyroid. When the free thyroid hormone level falls in the blood, the
negative-feedback effect of T3 on thyrotrophs is reduced, and the rate of TSH secretion
increases. The rise in TSH in the blood stimulates the thyroid gland to secrete thyroid
hormones at a greater rate. This action of T3 on thyrotrophs is thought to be due to changes
in gene expression in these cells.
TSH intervenes in all steps of Th secretion. It stimulates the activity of iodine pump,
proteolysis of thyroglobulin, followed by released of T3 and T4 in blood, iodination of tyrosin,
increase the number and activity of thyroid cells. Its effects are mediated by cAMP.
The life time of TSH = 60 minutes and its normal plasma level = 2 U/mL.
There are also other factors that are involved in regulation of Th secretion. The low
BMR, leptin and alpha melanocyte stimulating hormone (α MSH) increase the release of TRH
and synthesis of T4. The excess intake of iodine, stress, glucocorticoids, Dopamine,
Somatoststin, the low body temperature (only in infants) decrease the synthesis of Th by
inhibiting the secretion of TRH .

Disturbances of thyroid gland secretion

Hypothyroidism

If there is an insufficient amount of T3/T4, a condition referred to as hypothyroidism

results. Hypothyroidism leads to myxedema in adults and cretinism in children.
Symptoms of hypothyroidism derive from the fact that there is a reduction in the rate
of oxidative energy-releasing reactions within the body cells. Usually the patient shows puffy
skin, sluggishness, and lowered vitality. Other symptoms of hypothyroidism include weight
gain, decreased libido, inability to tolerate cold, muscle pain and spasm, insomnia and brittle
nails. Hypothyroidism in children, a condition known as cretinism, can result in mental
retardation, dwarfism, and permanent sexual immaturity. In severe hypothyroidism, a
substance consisting of hyaluronic acid and chondroitin sulfate complexed with protein is
deposited in the extracellular spaces of the skin, causing water to accumulate osmotically. This
effect gives a puffy appearance to the face, hands, and feet called myxedema.

Hyperthyroidism

Sometimes the thyroid gland produces too much thyroid hormones, a condition known
as hyperthyroidism. This condition produces symptoms such as an abnormally high body
temperature, profuse sweating, high blood pressure, loss of weight, irritability, and muscular
pain and weakness. It also causes the characteristic symptom of the eyeballs protruding from
the skull called exopthalmia. Hyperthyroidism has been treated by partial removal or by partial
radiation destruction of the gland. More recently, several drugs that inhibit thyroid activity
have been discovered, and their use is replacing the former surgical procedures.
Unfortunately thyroid conditions require lifetime treatment and because of the body's need
for a sensitive balance of thyroid hormone both supplementing and suppressing thyroid
function can take months or even years to regulate.

21
 In hyperthyroidism, if food intake is not increased as the metabolic rate is increased,
the body’s protein and fat stores are used up and the person loses weight. There is associated
muscle weakness, partly the result of the breakdown of muscle protein. However, the muscles
react more vigorously with increased hormones. Hyperthyroidism is associated with a fine
muscle tremor caused by the increased sensitivity of the nerves that control muscle tone. As
a result of the increase in metabolic rate, more heat is produced and there is a rise in body
temperature. This triggers the body mechanisms that regulate heat. The blood vessels in the
periphery dilate to dissipate heat, there is increased sweating, and the person becomes
intolerant to heat—all typical signs of hyperthyroidism. There is also a rise in heart rate and
cardiac output, partly as a result of its effect on increasing the sensitivity of the heart to
adrenaline and noradrenaline. The effect on metabolism raises the vitamin requirements;
vitamin deficiency is often associated with hyperthyroidism.
Although thyroid hormones do not increase the metabolism of the uterus, they are
required for normal functioning of the reproductive system. Menstrual irregularities are
associated with thyroid abnormalities. Excessive thyroid hormones make the person irritable
and restless. In short, the signs and symptoms of hyperthyroidism or thyrotoxicosis are that
of overactivity of the sympathetic nervous system—anxiety, nervousness, rapid heart rate,
sweating, tremor, and diarrhea. Sometimes the metabolic rate of the hyperthyroid individual
accelerates out of control and the person has a rapid heart rate, high fever, and other
symptoms of high metabolism. This situation is termed thyrotoxic crisis or thyroid storm and
requires immediate medical intervention.

Graves’ Disease

Graves’ disease is a thyroid gland disorder in which excessive thyroid hormones are
produced. Its cause is believed to be a result of autoimmunity - antibodies directed against
the TSH receptor in the plasma membranes of thyroid follicular cells. These antibodies (thyroid
stimulating immunoglobulin - TSI) bind to the TSH receptor, resulting in an increase in the
activity of adenylyl cyclase. The consequent rise in cAMP in follicular cells produces effects
similar to those caused by the action of TSH. The antibodies act for around 12 hours by
comparison with TSH that acts only for 1 hour. The high concentration of Th suppresses the
secretion of TSH.
The thyroid gland enlarges to form a diffuse toxic goiter, which synthesizes and
secretes thyroid hormones at an accelerated rate, causing thyroid hormones to be chronically
elevated in the blood. Feedback inhibition of thyroid hormone production by the thyroid
hormones is also lost. All hyperthyroidism symptoms, together with protrusion of the eyes
(exophthalmos) are observed. Exophthalmos may be caused by swelling of the eye muscles
within the bony orbit or inflammatory reactions caused by accumulation of immune
complexes connective tissue and fluid behind the eye.

PARATHYROID GLAND

Calcium and phosphorus in the organism

The amount of calcium in the organism is 1-2 kg. 99% of it is found in bones, and only
1% is in other compartments: 0.3% in muscles, 0.1% is in extracellular fluid, 0.01% is

22
intracellular fluid. [Ca2+] in circulation (calcemia) = 9.0 - 10.5 mg%. 40 - 42% of it is bound to
plasma proteins (it is not diffusible - it serves as a buffer of free plasma calcium), 60% is
diffusible (50% free or ionized and 8 -10% bound to small anions - citrate, phosphate, and
bicarbonate ).
There are 500 – 800 g of phosphorus in the body, most is in bones and teeth (85 -
90%). Compared with calcium, a much larger percentage of phosphorus is located in cells (10
-15%). The amount of phosphorus in extracellular fluid is low (0.08% of body content). In adult,
the normal range of plasma concentrations is 3.0 to 4.5 mg% (expressed in terms of milligrams
of phosphorus).
Phosphorus circulates in the plasma primarily as inorganic orthophosphate (PO4). At
a normal blood pH of 7.4, 80% of the phosphate is in the HPO42– form and 20% is in the H2PO4-
form. Nearly all plasma inorganic phosphate is ultrafilterable. In addition to free
orthophosphate, phosphate is present in small amounts in the plasma in organic form, such
as in hexose or lipid phosphates.
The absorption of Ca2+ takes place at level of intestine, excretion of calcium at level of
kidney. Daily 500 mg of calcium are mobilized, either stored or released from bones. Since
bone calcium serves as a reservoir, both bone resorption and bone formation are important
in regulating plasma calcium concentration. A small portion of this exchangeable calcium is
also the calcium found in all tissue cells, especially in highly permeable types of cells such as
those of the liver and the gastrointestinal tract. Most of the exchangeable calcium is in the
bone. It normally amounts to about 0.4 to 1% of the total bone calcium (10 g). This calcium is
deposited in the bones in a form of readily mobilizable salt (CaHPO4) and other amorphous
calcium salts. The importance of exchangeable calcium is that it provides a rapid buffering
mechanism to keep the calcium ion concentration in the extracellular fluids from rising to
excessive levels or falling to very low levels under transient conditions of excess or decreased
availability of calcium.

Absorption and excretion of calcium and phosphorus

Daily intake of calcium is around 1 g. Of this amount, only 1/3 is absorbed, the
remainder is removed through stool. An additional 250 mg/day of calcium enters the
intestines via secreted gastrointestinal juices and sloughed mucosal. The amount which is
absorbed, should be in balance with the amount which is excreted by the body. Thus, about
90 per cent (900 mg/day) of the daily intake of calcium is excreted in the feces.
Daily are filtered around 10 000 mg of calcium. 99% of the filtered calcium is reabs.
in the kidney (proximal (60%) , distal tubule (9%) and LH (30%) and only 1% of what is filtred
is excreted ( 100 - 150 mg/day = 10% of the ingested Ca ). Renal calcium excretion is controlled
primarily in the late distal tubule; parathyroid hormone stimulates calcium reabsorption here.
The absorption of calcium depends on the types of food ingested. There are some
substances in food that inhibite the absorption of calcium (phosphates, oxalates, found in
vegetables); also a diet rich in fats decreases the absorption of calcium; a diet rich in proteins
and lactose increases the absorption of calcium. The absorption takes place at the level of
duodenum and initial portion of jejunum and at the level of terminal ileum. At the level of
initial portion of small intestine is done by an active process, that has a maximum absorption
capacity. It means the rest is absorbed at the level of terminal ileum. At the terminal ileum the
absorption is done by diffusion.

23
 An adult ingests approximately 1,400 mg/day of phosphorus. In contrast to calcium,
most (1,300 mg/day) of this phosphorus is absorbed from the GI tract, typically as inorganic
phosphate. There is a secretion of phosphorus in the GI tract (about 200 mg/day), resulting
in a net uptake of phosphorus of 1,100 mg/day and excretion of 300 mg/day via the feces.
Thus, the majority of ingested phosphate is absorbed from the GI tract and little passes
through to the feces. Absorption of phosphorus occurs by active transport and passive
diffusion, but active transport is the primary mechanism. 80% or more of ingested phosphate
is absorbed.
About 85% of filtered phosphate is reabsorbed and 15% is excreted in the urine.
Phosphate reabsorption occurs by active transport, mainly in the proximal tubule where 65%
to 80% of filtered phosphate is reabsorbed. Parathyroid hormone inhibits phosphate
reabsorption in the proximal tubule and has a major regulatory effect on phosphate
homeostasis. It increases urinary phosphate excretion, leading to the condition of
phosphaturia. Calcitotnin also decreases plasma level of phosphate by inhibiting bone
resorption and stimulating urinary excretion.
The absorption of calcium depends on vitamin D3. Vitamin D3 stimulates the
synthesis of calcium-binding protein. This protein remains in the cells for several weeks after
the 1,25-dihydroxycholecalciferol (active form of vit. D3) has been removed from the body,
thus causing a prolonged effect on calcium absorption.
Vitamin D is found either in plants (Vitamin D2- ergocalciferol) or in animals. Vitamin
D3 (cholecalciferol) can be provided by the diet or formed in the skin by the action of
ultraviolet light on a precursor, 7-dehydrocholesterol, derived from cholesterol. In liver,
vitamin D3 is hydroxylated in position 25, giving birth to 25 hydroxycolecalciferol. At the level
of kidney, one more hydroxyle group at position 1 is introduced by a reaction catalyzed by an
enzyme named 1 alpha-hydroxylase. The activity of this enzyme is stimulated by parathyroid
hormone. When the concentration of calcium in blood is higher, then the activity of this
enzyme is inhibited. In this case is formed 24,25(OH)2dehydroxycolecalciferol which is
inactive.
Because the metabolits of vitamin D are synthesized in different organs and then
transported through blood to other organs , vitamin D3 can be considered as an hormone.

24
 (after A.Despopoulos, S. Silbernagl, Color Atlas of Physiology, 2003)

The receptor for Vitamin D3 is at the level of nucleus. Receptors for vitamin D3 are
found in bones, liver, placenta, brain, lymphocytes and parathyroid glands. At the level of
bones, vitamin D3 in small concentrations stimulates the mineralization of bone because it
activates the alkaline phosphatase from osteoblasts. In high concentration, vitamin D3
stimulates bone resorption. In high concentration it inhibites the secretion of parathyroid
hormone.
Vitamin D also promotes phosphate absorption by the small intestine. It is believed
that this results from a direct effect of 1,25-dihydroxycholecalciferol, but it is possible that it
results secondarily from this hormone’s action on calcium absorption. Calcium in turn acts as
a transport mediator for the phosphate.
Vitamin D also increases calcium (in the proximal part of collecting tubule) and
phosphate reabsorption by the epithelial cells of the renal tubules, thereby tending to
decrease excretion of these substances in the urine.
Vitamin D plays important roles in both bone absorption and bone deposition. The
administration of high quantities of vitamin D causes rebsorption of bone. In the absence of
vitamin D, the effect of PTH in causing bone resorption is greatly reduced or even prevented.
The mechanism of this action of vitamin D is not known, but it is believed to result from the
effect of 1,25-dihydroxycholecalciferol to increase calcium transport through cellular
membranes. Vitamin D in smaller quantities promotes bone calcification. One of the ways in
which it does this is to increase calcium and phosphate absorption from the intestines.
However, even in the absence of such increase, it enhances the mineralization of bone.
The net effect of 1,25-dihydroxycholecalciferol is to increase both calcium and
phosphate concentrations in plasma. The activated form of vitamin D primarily influences the

25
GI tract, although it has actions in the kidneys and bones as well. In the kidneys, 1,25-
ihydroxycholecalciferol increases the tubular reabsorption of calcium and phosphate,
promoting the retention of both ions in the body. However, this is a weak and probably only
minor effect of the vitamin.

Physiology of bones

2/3 of bones are made up by mineral substances and 1/3 by organic substances.
Mineral substances are represented by calcium and phosphates, bound together, forming
hydroxyapatite crystals [Ca10(PO4)6(OH)2]. Bone contains also a lot of magnesium,
carbonates, sodium, potassium. Bones may take some radioactive substances such as:
strontium, plutonium, uranium, heavy metals (lead). After exposition to radiations, malignant
osteonoma can occur.
Organic matrix is made up 95% by collagen fibers (type I collagen) and 5% of ground
substance. The ground substance is composed of extracellular fluid plus proteoglycans,
especially chondroitin sulfate and hyaluronic acid. Organic matrix is also called osteoid. It is
impregnated by hydroxyapatite cristals that give the bones a powerful structure. Electron
microscopic study of bone reveals needle-like hydroxyapatite crystals lying alongside collagen
fibers. This orderly association of hydroxyapatite crystals with the collagen fibers is
responsible for the strength and hardness characteristic of bone.
At the level of bones 3 types of cells can be found. Osteoblasts are very active
metabolic cells having a diameter around 20 micrometer. They have a rich endoplasmic
reticulum and ribosomes. They secrete collagen. As long as osteoblast secrete the osteoid, it
becomes mineralized, then osteoblasts slowly transform into osteocytes. Osteoblasts and
osteocytes have processes. These processes are found inside of some canalicules. By these
they communicate with the surface of bones, get nutritive substances and at the same time
waste product of their metabolisms are taken away. PTH produces an enlargement of spaces
surrounding osteocytes.
Osteoclast is another type of cell. They are the macrophages of bone tissue, which
derive from monocytes. They are multinucleated cells, having the role to erode the structure
of bone. They contain acid phosphatase and proteolytic enzymes by which osteoid is broken
down, and also some acidic substances like citric acid and lactic acid by which calcium salts
are removed from this structure. Bone resorption is a two-step process. First, osteoclasts
create a local acidic environment that increases the solubility of surface bone mineral. Second,
proteolytic enzymes secreted by osteoclasts degrade the organic matrix of bone.
In order that calcium and phosphate to form hydroxyapatite, it is necessary that
calcium and phosphate to be in a certain concentration to become saturated and precipitate.
Alkaline phosphatase released by osteoblast have the role to hydrolyze phosphoric esters and
to supply the concentration of phosphates to the proximity of osteoblasts in order that
calcium and phosphate to precipitate and to store as hydroxyapatite. The concentration of
calcium and phosphate ions in extracellular fluid are considerably greater than those required
to cause precipitation of hydroxyapatite. However, inhibitors are present in almost all tissues
of the body as well as in plasma to prevent such precipitation; one such inhibitor is
pyrophosphate. Therefore, hydroxyapatite crystals fail to precipitate in normal tissues except
in bone despite the state of supersaturation of the ions.

26
 Parathyroid glands

They are 4 small glands placed on the posterior side of the thyroid gland. These glands
are vital and they weight together 120 mg. Parathyroid glands are made up of 2 types of cells:
1) chief cells having a clear cytoplasm. They secrete Parathyroid hormone (PTH); 2) oxyphils
cells with many granules in their cytoplasm. Their function is not very well known - they are
believed to be modified or depleted chief cells that no longer secrete hormone.
Parathyroid hormone is a polypeptide made up of 84 aa, having a molecular weight of
9500 D. It is secreted as a preprohormone of 110 aa, that is processed in ER and Golgi
apparaturs and stored in vesicles. Parathyroid hormone has the receptor at the level of cell
membrane, which is coupled with G protein; it mediates its activity by adenylyl cyclase/cAMP.
Its plasma concentration = 1.5 – 5.5 ng/dL. 60 –70% of PTH is degraded by Kupffer cells in liver
(by proteolysis) and 20 - 30% is degraded in kidneys and less in other organs. Parathyroid
hormone has several roles:
- it stimulates the absorption of calcium at the level of small intestine. It does this
indirectly, because PTH activates 1- alpha hydroxylase at the level of kidney, necessary for
formation of active vitamin D3.
- it stimulates the resorption of bones. PTH increases the permeability of osteoblast
and osteocyte membrane to calcium. It stimulates the activity of calcium pump from the
membrane of these cells, so calcium moves faster into blood. This process occurs within
minutes from its release into the circulation and explains the rapid phase of rising calcemia.
In the second phase (slow phase) osteoclasts are activated, resulting in the resorption of bone
and moving of calcium into blood.
In addition to stimulate, PTH induces the maturation of immature osteoclasts into
mature, active osteoclasts. PTH inhibits the synthesis of collagen by osteoblast. The actions of
PTH to promote bone resorption are augmented by 1,25-dihydroxycholecalciferol. PTH
increases the urinary excretion of hydroxyl-proline. PTH by means of cyclic AMP stimulates
the synthesis and release of lysosomal enzyme that contributes to dissolution/resorption of
organic matrix. Histological and physiologic studies have shown that PTH causes removal of
bone salts from two areas in the bone: (1) from the bone matrix in the vicinity of the
osteocytes lying within the bone itself and (2) in the vicinity of the osteoblasts along the bone
surface.
- PTH increases the reabsorption of calcium at the level of thick ascending limb and
distal tubule. The amount of calcium excreted daily is less than 1% of the amount which is
filtered. At the same time PTH inhibits the reabsorption of phosphates at the level of proximal
tubule.
- it increases the rate of reabsorption of magnesium ions and hydrogen ions while it
decreases the reabsorption of sodium, potassium, and amino acid ions in the same way that
it affects phosphate.
The secretion of PTH is not under the control of pituitary gland. When the
concentration of calcium (ionized) decreases below 9.5 mg%, the secretion of PTH is
stimulated, while the increase of calcium concentration inhibits the secretion of PTH.

Calcitonin (CT)

It is a polypeptide containing 32 aa, (MW = 3500 D), whose main role is to reduce the
concentration of calcium in blood, but it also reduces the concentration of phosphates. Plasma

27
conc. of CT = 1-2 ng/dL and its life time = 5 – 10 min. Calcitonin acts by impeding the resorption
of bone. Calcitonin stimulates the activity of osteoblasts, and inhibits the activity of
osteoclasts.
CT secretion is stimulated by an increase in plasma calcium concentration. Hormones
of the GI tract, especially gastrin, also promote CT secretion. Because the net effect of CT is to
promote calcium deposition in bone, the stimulation of CT secretion by GI hormones provides
an additional mechanism for facilitating the uptake of calcium into bone after the ingestion of
a meal. The regulation of calcitonin secretion is not under the control of pituitary gland. It is
done by means of level of calcemia.
The overall action of calcitonin is to decrease both calcium and phosphate
concentrations in plasma. The primary target of CT is bone, although some lesser effects also
occur in the kidneys. In the kidneys, CT decreases the tubular reabsorption of calcium and
phosphate. This leads to an increase in urinary excretion of both calcium and phosphate and,
ultimately, to decrease the level of both ions in the plasma. In bones, CT opposes the action
of PTH on osteoclasts by inhibiting their activity. This leads to decreased bone resorption and
an overall net transfer of calcium from plasma into bone. CT stimulates osteoblastic activity
and deposition of calciumnad phosphate in bones. Calcitonin prevents the absorption from
intestine.
The secretion of this hormone is stimulated when calcemia increases above 9.5 mg%.
The secretion is also stimulated by beta adrenergic substances (dopamine, epinephrine,
norepinephrine), glucagon, cholecystokinin. The concentration of calcitonin increases in
pregnancy and by this it is considered that it protects the mother bones from being
demineralized.

Disturbances of parathyroid gland

Hypofunction

When calcemia decreases below 7 mg%, then spasmophilia or hypocalcemic tetania

occurs, which is characterized by spontaneous contraction of muscles and spasms at the level
of larynx muscles. In this case there is an increased excitability. If calcium concentration
decreases below 5 mg% = tetany.
When the extracellular fluid concentration of calcium ions falls below normal, the
nervous system becomes progressively more excitable, because this causes increased
neuronal membrane permeability to sodium ions, allowing easy initiation of action potentials.
At plasma calcium ion concentrations about 50 per cent below normal, the peripheral nerve
fibers become so excitable that they begin to discharge spontaneously, initiating trains of
nerve impulses that pass to the peripheral skeletal muscles to elicit tetanic muscle contraction.
Osteosclerosis is secondary to intoxication with Pb or to some bone tumors. In this
case there is an extra amount of bone tissue. Osteoclasts lost their capacity to resorb bone,
and by this there is a decrease of bone cavities, of bone marrow, and also a narrowing of
orifices by which nerves penetrate bones.
Osteoporosis - there is a decrease of bone mass. The structure of bone becomes
more porous, more fragile, and there is an increased risk of fracture. Osteoporosis involves a
reduction in total bone mass with an equal loss of both bone mineral and organic matrix.

28
Because the activity of osteoblasts is under the control of oestrogen, at menopause, the
oestrogen secretion stops. This may result in osteoporosis.

Parathyroid hyperfunction

Hypercalcemia is a condition which is noted when blood calcium level is above 12

mg/dL. Its common signs are: depression of the NS, lack of appetite, constipation, sluggishness
of reflexes, reduced ST segm. and QT interval on EKG, bone diseases.
The cystic bone disease of hyperparathyroidism is called osteitis fibrosa cystica.
Osteoblastic activity in the bones increases greatly in an attempt to form enough new bone
to make up for the old bone absorbed by the osteoclastic activity. When the osteoblasts
become active, they secrete large quantities of alkaline phosphatase. Therefore, one of the
important diagnostic findings in hyperparathyroidism is a high level of plasma alkaline
phosphatase. Bone resorption is increased and in bones can be formed some kyst (cavities),
that is why it is named fibrosa cystica.

THE ENDOCRINE PANCREAS

It is represented by Langerhans islets. They are the structural unit of the endocrine
pancreas. There are between 1-2 millions islets in pancreas. They account for 1-2% of the
weight of pancreas, around 1 g. They are found in the tail and less in the head. Each islet is
formed by a few hundreds to a few thousands of cells. There are 4 types of cells, producing 4
hormones. In the middle of these islets there are beta cells: 60 -70 –90 % of the cells are beta
cells (diam= 10 –15 microm), they produce insulin and amylin. 20 - 25% of cells are alpha cells
producing glucagon, they are found at the periphery of the islets. Between 1-8% (at the
periphery) of cells are delta cells, producing somatostatin. Less than 2% are F cells, they
produce pancreatic polypeptide.

Insulin

MW = 6000 D, 51 aa. It is secreted as a preprohormone at the level of endoplasmic

reticulum of beta cells. The genes for synthesis of insulin are localized on pair 11 of
chromosome. In ribosomes is produced preproinsulin, which is transferred to the endoplasmic
reticulum, where it loses 23 aa, becoming pro-insulin. Proinsulin is made up of a peptide A
having 21 aa, a peptide B made up of 30 aa, and a connecting peptide between these two
named peptide C, made up of 31 aa. Peptide C is necessary for the petide A and B to have a
certain position in space in order to form sulfidic bridges. Peptide C is removed under the
action of an enzyme (trypsin-like) and is transformed into insulin. After that insulin is packed
in membranes at the level of Golgi apparatus and is stored in some granules at the level of the
cell.
Insulin is released from beta cell by exocytosis, which is a calcium dependent
phenomenon. In blood, insulin is secreted together with peptide C in equimolar amounts, but
peptide C does not have a biological activity. Also small amounts of proinsulin are secreted in

29
blood. Proinsulin has only 10% of the activity of insulin, but it cannot be transformed into
insulin outside of the pancreas. Insulin circulates in blood almost entirely unbound.
Peptide C is used in clinic in order to monitorize the secretory activity of the endocrine
pancreas in people suffering from diabetes mellitus, who receive exogenous insulin.
Pancreas secretes daily between 40-60 units of insulin, but in pancreas there is a pool
of 200 units of insulin.
Based on studies using isolated animal pancreas preparations maintained in vitro, it
has been determined that insulin is secreted in a biphasic manner in response to a marked
increase in blood glucose. An initial burst of insulin secretion may last 5 to 15 minutes,
resulting from the secretion of preformed insulin secretory granules. This response is followed
by more gradual and sustained insulin secretion that results largely from the synthesis of new
insulin molecules. It reaches the maximum between 2 – 2.5 hours.
After insulin is released into blood, it is taken by several organs having receptors for
insulin - liver, muscles and fat tissue. Brain and RBCs do not uptake insulin, that means the
penetration of glucose inside neurons and RBCs is independently on insulin. Except for that
portion of the insulin that combines with receptors in the target cells, the remainder is
degraded by the enzyme insulinase mainly in the liver, to a lesser extent in the kidneys and
muscles, and slightly in most other tissues.
The lifetime of insulin is 5 minutes. Normal concentration of insulin (a jeun- basal
concentration) in blood = 6 – 26 µU/mL.

Effects of Insulin

The main effect is to decrease glycemia. Normal glycemia is 70 -115 mg/dL.

Intravenous administrated insulin decreases glycemia, which reaches the lowest value within
30 minutes. If it is administrated subcutaneously, the lowest value is reached in 2 to 4 hours.
The receptor of insulin (MW= 340,000 D) is made up of 2 alpha sub-units and 2 beta
sub-units connected by sulfuric bridges. Alpha-subunits are extracellular, and beta subunits
cross the cell membrane. The intracellular surfaces of beta subunits have the tyrosine-kinase
activity. A receptor can bind 2 molecules of insulin. After reaction with insulin, there is a
conformational change of R and beta subunits are activated. Activation of the beta subunit of
the insulin receptor results in autophosphorylation, involving the phosphorylation of a few
selected tyrosine residues in the intracellular portion of the receptor. This event further
activates the tyrosine kinase portion of the beta subunit, leading to tyrosine phosphorylation
of specific intracellular substrates. A cascade of events follows, leading to the pleiotropic
actions of insulin in its target cells. While tyrosine phosphorylation events appear to be the
early steps in insulin action, serine/threonine phosphorylation or dephosphorylation is
involved in many of the final steps of insulin action.
The number of receptors for insulin on the cell is variable. When the concentration
of insulin increases, the number of receptors on the cells decreases. In people suffering from
obesity, the number of insulin R lowers, and in this way is explained their resistance to insulin.
During starvation, the number of receptors for insulin increases, because the
concentration of insulin decreases. When there is an overfeeding, the secretion of insulin is
stimulated continuously, so the number of receptors will decrease, and this process will
produce a resistance to insulin.
Insulin is an anabolic hormone. Its main role is to decrease the concentration of
glucose in circulation. It is the only hormone that decreases the concentration of glucose in

30
blood. Insulin performs this effect by several mechanisms. Insulin stimulates the uptake of
glucose in different cells (facilitated diffusion), especially in the liver, muscle and adipose
tissue, mammary glands, bones, connective tissue, WBCs, peripheral nerves, alpha cells of
pancreas. Most of the cells of the organism have receptors for insulin.
There are a few tissues where glucose can penetrate without insulin. These tissues
are: brain (except hypothalamus), RBCs, enterocytes, renal tubular cells and beta cells of
pancreas. After the ingestion of foods, glucose is absorbed and under the action of insulin, the
penetration of glucose inside the hepatocytes is stimulated.
Considerable recent work has revealed not just one transporter, but a family of about
seven different glucose transporters (GLUT), commonly called GLUT 1 to GLUT 7. These
transporters are expressed in different tissues and, in some cases, at different times during
fetal development.
GLUT 4, the insulin-stimulated glucose transporter, is the primary form of the
transporter present in skeletal muscle tissue and adipose tissue. It is present in plasma
membranes and in intracellular vesicles of the smooth ER. In target cells, the effect of insulin
is to promote the translocation of GLUT 4 transporters from the intracellular pool into plasma
membranes. As a result, more transporters are available in the plasma membrane, and
glucose uptake by target cells is, thereby, increased.
Glucose is stored in hepatocytes as glycogen. In this way, a gradient of concentration
for glucose is maintained in order that glucose continues penetrating inside of hepatocytes.
Glucose can be stored as glycogen until glycogen reaches the value 5-6% of liver’s mass. During
inter-digestive periods, glycogen is broken down and glucose is released. In this way is
maintained a certain level of glycemia, even in the periods when the person does not eat
(starvation period). A constantly supply of energy is assured.
Under the action of insulin, glycogen synthesis is stimulated. This is due because
insulin inhibits the breakdown of glycogen. Insulin inhibits glycogen phosphorylase, so
glycogenolysis is inhibited. Glucokinase is activated, and glucose is phosphorylated, being
transformed into glucose-6-phosphate. G6P is a product that cannot leave the hepatocytes.
At the same time, other enzymes involved in glycogen synthesis are stimulated by insulin:
phosphofructo kinase and glycogen synthase. The amount of glucose that cannot be stored in
hepatocytes as glycogen is transformed into fatty acid, and they will be transported and stored
as triglycerides at the level of adipose tissue.
Insulin also enhances glycolysis - in addition to increasing glucose uptake and
providing a mass action stimulus for glycolysis, insulin activates the enzymes glucokinase,
hexokinase, phosphofructokinase, pyruvate kinase, and pyruvate dehydrogenase of the
glycolytic pathway.
Muscles cannot take glucose in the absence of insulin, so in resting condition, they
cannot uptake glucose. That’s why in resting conditions, they use specially fatty acids as
energetic substrates for the synthesis of ATP, but after a meal, when the secretion of insulin
is stimulated, glucose can penetrate in striated muscles by facilitated diffusion. At the same
time glucose can be stored in muscles as glycogen, but the amount stored in muscle is lesser
than the amount stored in the liver. Glycogen can be stored till the level reaches 1% (or 2 –
3%) of the muscle mass. Glycogen stored in muscle is used specially to supply energy in
anaerobic conditions. In muscles being in activity, glucose can penetrate even without insulin,
because sarcolemma becomes more permeable to glucose during physical activity. This is why
the dose of insulin should be reduced in people with diabetes making a physical effort.

31
 Insulin inhibits gluconeogenesis. It does this mainly by decreasing the quantities and
activities of the liver enzymes required for gluconeogenesis. However, part of the effect is
caused by an action of insulin that decreases the release of amino acids from muscle and other
extrahepatic tissues and in turn the availability of these necessary precursors required for
gluconeogenesis.
Insulin increases the synthesis of FA (fatty acids) in the liver. Glucose that is not
stored as glycogen (in liver) is transformed in acetyl-CoA, and acetyl-CoA in fatty acids. Fatty
acids are transported as TG in lipoproteins (VLDL) in fatty tissue and they are stored in adipose
tissue as TG. Glucose can also penetrate in adipose tissue, but under the action of insulin.
Glucose is broken down by glycolysis, and by this process great amount of alpha-
glycerolphosphate is formed. From alpha-glycerophosphate, glycerol is released, that reacts
with fatty acids forming triglycerides. At the same time insulin at the level of adipose tissue
inhibits a hormone dependant lipase and by this triglycerides are not broken down.
Mobilisation of lipids from adipose tissue is inhibited.
In addition to promoting de novo fatty acid synthesis in adipose tissue, insulin
increases the activity of lipoprotein lipase, which plays a role in the uptake of fatty acids from
the blood into adipose tissue. As a result, lipoproteins synthesized in the liver are taken up by
adipose tissue, and fatty acids are ultimately stored as triglycerides.
Insulin inhibits lipolysis, that is why in the absence of insulin, or decrease amount of
insulin, like in diabetes mellitus, this lipase will not be inhibited. Triglycerides will be broken
down, fatty acids will be released, they will be taken by liver and at level of mitochondria in
hepatocytes, these fatty acids will be oxidized forming acetyl-CoA. A certain amount of acetyl-
CoA can be used by hepatocytes as energetic source, but most of acetyl-CoA condensates
forming acetoacetic acid that can pass into circulation. In healthy persons, most of the
peripheral cells transform acetoacetic acid into acetyl-CoA. But when there is a deficit of
insulin, this transformation is not produced, and a part of it is transformed into beta
hydroxybutyric acid and acetone (ketone bodies). Their acculumation (of ketone bodies) can
lead to a state of ketosis and acidosis, and even coma.

Effects of insulin on lipid metabolism in adipocytes (Rhoades R, Tanner G, Medical Physiology,

second edition, Lippincott Williams & Wilkins, 2003)

32
 Insulin stimulates the transport of amino acids in cells, and at the same the
synthesis of proteins in ribosomes. Insulin actually increases the permeability of cell mb. for
aa. Protein anabolism is stimulated. Catabolism of proteins is inhibites by Insulin. In liver,
insulin inhibits the synthesis of glycogenogenetic enzymes, so preventing the release of aa
from muscle and other extrahepatic tissues.
In diabetes when there is less insulin, catabolism of protein is stimulated which leads
to a negative nitrogen balance. Because proteins are not synthesized properly, also antibodies
are not synthesized, which explains the tendancy towards infection in people suffering form
diabetes. At the same time because of high concentration of glucose in diabetes, the growing
of germs is stimulated. Glucose from the fluids of the organism is a good medium for growing
of bacteria.
Insulin stimulates the transport of potassium, phosphate and calcium inside of cells.
This is principally due to the stimulation of the activity of sodium potassium ATPase. A side
effect of high doses of insulin is hypopotassemia. Decrease of plasma level of K + in its turn
lowers the secretion of insulin. Thiazidic diuretics, that induce loss of Na + and K+ through the
urine, may reduce the tolerance to glucose and aggravates diabetes.
Along with GH, insulin promotes growth of body.

Regulation of insulin secretion

The main factor stimulating the secretion of insulin is the level of glycemia. Anytime
the concentration of glucose in blood increases, the secretion of insulin is stimulated.
Secretion of insulin takes place in the following manner: glucose penetrates inside of beta
cells. Once inside the cells, glucose is phosphorylated to glucose-6-phosphate by glucokinase.
This step appears to be the rate limiting for glucose metabolism in the beta cell and is
considered the major mechanism for glucose sensing and adjustment of the amount of
secreted insulin to the blood glucose levels. The glucose-6-phosphate is subsequently oxidized
to form adenosine triphosphate (ATP), which inhibits the ATP-sensitive potassium channels of
the cell. Closure of the potassium channels depolarizes the cell membrane, thereby opening
voltage-gated calcium channels, which are sensitive to changes in membrane voltage. This
produces an influx of calcium that stimulates fusion of the docked insulin-containing vesicles
with the cell membrane and secretion of insulin into the extracellular fluid by exocytosis. The
secretion of insulin increases when the concentration of glucose is about 100 mg%, and the
latency is between 30-60 seconds.
Other nutrients, such as certain amino acids, can also be metabolized by the beta cells
to increase intracellular ATP levels and stimulate insulin secretion. Some hormones, such as
glucagon and gastric inhibitory peptide, as well as acetylcholine increase intracellular calcium
levels through other signaling pathways and enhance the effect of glucose, although they do
not have major effects on insulin secretion in the absence of glucose. Other hormones,
including somatostatin and norepinephrine (by activating adrenergic receptors), inhibit
exocytosis of insulin.
At the normal fasting level of blood glucose, the rate of insulin secretion is minimal—
on the order of 25 ng/min/kg of body weight. If the blood glucose concentration is suddenly
increased to a level two to three times normal and kept at this high level thereafter, insulin
secretion increases markedly in two stages. Plasma insulin concentration increases almost 10-
fold within 3 to 5 minutes after the acute elevation of the blood glucose; this results from

33
immediate dumping of preformed insulin from the beta cells of the islets of Langerhans. The
initial high rate of secretion is not maintained; instead, the insulin concentration decreases
about halfway back toward normal in another 5 to 10 minutes. Beginning at about 15 minutes,
insulin secretion rises a second time and reaches a new plateau in 2 to 3 hours, this time
usually at a rate of secretion even greater than that in the initial phase. This secretion results
both from additional release of preformed insulin and from activation of the enzyme system
that synthesizes and releases new insulin from the cells.
Increasing the concentration of some amino acids (arginine, lysin, leucine), but also
ketone bodies stimulate the secretion of insulin. Adrenergic substances act on beta
receptors, also glucagon stimulate the secretion of insulin. Stimulation of SNS inhibits the
secretion of insulin by a direct action on pancreas. Parasynpathetic nervous system by means
of acetylcholine stimulates the secretion of insulin.
Stimulation of sympathetic nervous system by means of alpha receptors will inhibit
the secretion of insulin. [Alpha (inhibitory) and beta (stimulatory) adrenergic receptors are
located on beta cells of Langerhans islets]. Cholecystokinin, secretin, gastric inhibitory
peptide, enteroglucagon, stimulate directly the secretion of insulin. This explains why the
secretion of insulin is higher when the glucose is administrated orally by comparision with the
concentration of insulin that is lower when glucose is administrated through a perfusion.
Other hormones such as growth hormone stimulate the secretion of insulin,
adrenocorticotrope hormone (ACTH), and glucocorticoids by inducing hyperglycemia
stimulate the secretion of insulin. Thyroid hormones by increasing the absorption of glucose
at the level of digestive tract and by mobilizing hepatic glycogen, induce hyperglycemia and
induce secretion of insulin. Leptin, somatostatin and fasting decrease insulin secretion.
When the glucose concentration is low, insulin secretion is suppressed and fat is used
almost exclusively for energy everywhere except in the brain. When the glucose concentration
is high, insulin secretion is stimulated and carbohydrate is used instead of fat, and the excess
blood glucose is stored in the form of liver glycogen, liver fat, and muscle glycogen. Therefore,
one of the most important functional roles of insulin in the body is to control which of these
two foods from moment to moment will be used by the cells for energy.

Disturbance of insulin secretion

Diabetes mellitus

It can be induced experimentally in animals by removing of pancreas or by

administration of some drugs such as ALOXAN or STREPTOZOTOCIN. These drugs are
pancreato-toxic, they will destroy the beta cells. Diabetes mellitus according to the needs of
insulin, is classified in type I, also called insulin-dependant, and diabetes mellitus type II, also
called non insulindependant.
In case of type I, the pancreas is almost completely destroyed by an immune
mechanism. In case of type II diabetes, it occurs mostly in obese people. In this case there is
a decrease of secretion of insulin. It is considered that the decrease is the result of the
overfeeding. In obese people the secretion of insulin is constantly stimulated, and at a certain
moment the pancreas won’t be able to provide enough insulin and its secretion will be
reduced.

34
 Diabetes mellitus is a desease having an increasing incidence. In diabetes mellitus
there is hyperglycemia and an increased concentration of lipids. Because glucose cannot
penetrate in cells in the absence of insulin, the person will have a constant desire to eat
(polyphagia), and also glucose is lost through the urine (glucosuria). There is an osmotic
diuresis (polyuria), followed by dehydration and compensatory by an increased ingestion of
water (polydipsia). The main complications of diabetes are: atherosclerosis (myocardial
infarction, stroke), diabetic retinopathy, neuropathy and nephropathy.

Hyperinsulinism

It occurs when insulin is secreted in higher amounts and this is due to some tumors
at the level of pancreas. When glycemia decreases below 50 mg%, that can induce
hypoglycemic coma. When the level is between 50-70 mg% - tremors, sweating, tachycardia,
increased blood pressure, dizziness, speaking disturbances. This is due to the lack of glucose
at the level of the neurons and to the stimulation of sympathetic nervous system
(compensatory).

Glucagon (G)

Peptide made up of 29 amino acids, MW = 3500 D, half-life = 3-6 minutes. Glucagon

is synthesized from the precursor called preproglucagon in alpha cells of pencreas. 30% of G
ias degraded by liver, 20% by kidney and 50% of the circulating G is degraded in blood itself
by the enzymes ( serine and cysteine proteases).
It has the opposite effects to insulin. It induces hyperglycemia. The receptor for
glucagon is coupled with G-protein and it induces its effects by stimulation of adenyl cyclase
and increasing the concentration of AMPc. The AMPc will activate the enzyme involved in
glycogenolysis and gluconeogenesis. Glucagon stimulates glycogenolysis in the liver cells (it
activates phosphorilase and inhibits glycogen synthase), but not in muscle cells.
Gluconeogenesis: the main source - amino acids, but in order to be transformed into
glucose they need to be deaminated. That’s why this process is associated with increase or
ureogenesis. It does this principally by increasing the transcription of mRNA coding for the
enzyme phosphoenolpyruvate carboxykinase (PEPCK), a key rate-limiting enzyme in
gluconeogenesis. G increases the transport of aa into liver cells, that will be used for
gluconeogenesis.
At the level of adipose tissue, glucagon stimulates lypolysis. It has a catabolic role by
the fact it stimulates the mobilization of fatty acids, amino acids and glucose from the
storages. Glucagon promotes ketogenesis, the production of ketones, by lowering the levels
of malonyl CoA, relieving an inhibition of palmitoyl transferase and allowing fatty acids to
enter the mitochondria for oxidation to ketones. Ketones are an important source of fuel for
muscle cells and heart cells during times of starvation, sparing blood glucose for other tissues
that are obligate glucose users, such as the central nervous system.
Glucagon stimulates the secretion of insulin by acting on beta cells, but also the
secretion of growth hormone, somatostatin, and catecholamines.
In higher concentration than normal, glucagon increases the force of contraction of
heart. It also stimulates the biliary secretion and inhibits the secretion of gastric juice in
stomach.

35
 Regulation of glucagon secretion is done by the level of glycemia. When level of
glycemia decreases below 70 mg%, the secretion of glucagon is stimulated directly, but also
indirectly by activation of SNS. Excitation of SNS rises the secretion of glucagon by interaction
with alpha and beta R from the alpha cells of pancreas. Stimulation of parasympathetic
nervous system can also increase glucagon secretion.
A diet rich in proteins and amino acids (alanine, arginine) increases glucagon
secretion. Cholecystokinin and gastrin stimulate glucagon secretion, while insulin,
somatosatin inhibit glucagon secretion. A high level of exogenous or endogenous FFA
suppresses the release of glucagon from pancreas. During starvation, stress, physical effort
the plasma level of glucagon rises, while insulin level lowers.

Somatostatin

It is synthesized under two forms- 14 aa and 28 aa. It is produced by delta cells of

pancreas, but it is also synthesized in hypothalamus, spinal cord, stomach and upper part of
small intestin. . Its life time in circulation = 3 minutes. Somatostatin inhibits the secretion of
insulin, glucagon and pancreatic polypeptide. Somatostatin is first synthesized as a larger
peptide precursor, preprosomatostatin. Upon insertion of preprosomatostatin into the rough
ER, it is initially cleaved and converted to prosomatostatin. The prohormone is converted into
active hormone during packaging and processing in the Golgi apparatus.
Factors that stimulate pancreatic somatostatin secretion include hyperglycemia,
glucagon, and amino acids. Glucose and glucagon are generally considered the most important
regulators of somatostatin secretion. Somatostatin inhibits the secretion of growth hormone
and TSH at the level of pituitary gland.
It decreases the motility of digestive tract, reduces gastric and biliary secretion and
motricity of gall bladder. It reduces the secretion of GI hormones: gastrin, CCK, GIP, VIP.
Its secretion is inhibited by fatty acids. High levels of exogenous insulin inhibits the
release in the circulation of somatostatin, while glucagons intensifies it. It has been suggested
that the principal role of somatostatin is to extend the period of time over which the food
nutrients are assimilated into the blood. At the same time, the effect of somatostatin to
depress insulin and glucagon secretion decreases the utilization of the absorbed nutrients by
the tissues, thus preventing rapid exhaustion of the food and therefore making it available
over a longer period of time.

Pancreatic polypeptide

Made up of 36 aa, this hormone has a not well known role. It is supposed to increase
glucagon secretion from alpha cells, to reduce the exocrine secretion of pancreas, together
with decrease of absorption of food. Its secretion is under the control of PNS.
The secretion of this hormone has been noted to increase after hypoglycemia,
ingestion of proteins and starvation. Decrease of its secretion has been noted after
intravenous administration of glucose and by somatostatin.

ADRENAL GLAND

36
 There are two adrenal glands, located on the superior poles of the kidneys.Each gland
weighs about 4g. These glands have a vital role because if they are removed, this leads to
collapse and death. Adrenal gland is made up of cortex and medulla. Cortex represents 70-
90% of the gland, the remainder is medulla. At the level of cortex are described several layers:
zona glomerulosa, fasciculata, and reticullaris. The medulla secretes catcholamines. The
gland is surrounded by a capsule. Under the capsule there is zona glomerulosa (15% of gland),
which synthesizes some hormones named mineralocorticoids (aldosteron). Zone glomerulosa
is above the zona fasciculate (75%) which is made up of cords of cells. Between these cord of
cells there are fenestrated capillaries. Zona fasciculata synthesizes cortisol, corticosterone,
small amounts of adrenal androgens and estrogen. Beneath zona fasciculate, there is zone
retucularis, made of a network of cells. Zona fasciculata and reticularis secrete sex hormones
and glucorticoid hormones. The zona reticularis, the deep layer of the cortex, secretes the
adrenal androgens dehydroepiandrosterone (DHEA) and androstenedione, as well as small
amounts of estrogens and some glucocorticoids. The distinction between zona fasciculata and
reticularis is not very clear and they seem to function as a unit.

Adrenal gland ( after A. Despopoulos, S. Silbernagl, Color Atlas of Physiology, 2003)

The activity of adrenal gland is under the action of ACTH produced by pituitary gland.
It stimulates the synthesis of adrenal gland hormones and the normal functioning and

37
structure of the gland. Zona glomerulosa has a regenerative role. The other two zones can be
regenerated by zona glomerulosa. When zone glomerulosa is destroyed, then the structure of
adrenal gland cannot be recovered. The activity of zone glomerulosa is not under the control
of ACTH (or less influenced), but under angiotensin II.

Synthesis of adrenal gland hormones

Adrenal hormones are synthesized from cholesterol (these are steroids hormones).
They are not hydrosoluble. Most of used cholesterol comes from blood, from LDL, and to a
lesser extent it is synthesized from acetate inside of adrenhal gland cells. At the level of cell
mb. of adrenal cells there are receptors for LDL.
Adrenal gland synthesises two types of steroid hormones: 21 steroids and 19 steroids.
C 21 steroids are called in this way because there is attached a chain of carbon atoms in the
position 17. Mineralocorticoids and glucocorticoids are C21 steroid hormones.
While for the other category (C 19 steroids), at the level of position 17 there is a Keto
group or a hydroxyl group. C19 are sex hormones. The hormones having a Keto group in the
position 17 are also called Keto steroids.
Adrenocorticotrope hormone (ACTH) stimulates the uptake of cholesterol inside of
adrenal cells. Cholesterol is esterified and stored inside of cytoplasma as some droplets of
lipid.

Sources of cholesterol for steroid biosynthesis by the adrenal cortex (Rhoades R, Tanner G,
Medical Physiology, second edition, Lippincott Williams & Wilkins, 2003)

Under the action of a hydrolase, from the esterified cholesterol, free cholesterol is
removed. Free cholesterol is used in mytochondrion to produce pregnenolone (under the
action of a cholesterol–desmolase). Pregnenolone is transformed into progesterone under
the action of a dehydrogenase . Progesterone gives birth to aldosterone in zone glomerulosa,
and to cortisol and sex hormones (dehydroepiandrosterone) in zona fasciculata and zona
reticularis.

38
 The transformation of cholesterol into pregnenolone is stimulated by ACTH. It
mediates its action by mean of cAMP. cAMP activates a protein kinase A which will
phosphorylate different enzymes involved in conversion of esterified cholesterol into free
cholesterol and conversion of cholesterol into pregnenolone.

Transport of adrenal hormones in circulation and their metabolism

These hormones are not hydrosoluble. They are transported in plasma bound by
proteins. Glucocorticoid hormones (Cortisol is the main) are transported 90 - 95% by an alpha
2 globulin synthesized by liver named transcortin or cortisol-binding globulin. The remainder
is free in plasma. Albumin plays very little role in glucocorticoid transport. The other
glucocorticoid hormones, corticosterone, is also transported by transcortin and albumin.
Cortisol has a life time of 60 – 90 minutes (because most of it is bound in the circulation). Only
about 60% of circulating aldosterone combines with the plasma proteins, so that about 40%
is in the free form; as a result, aldosterone has a relatively short half-life of about 20 minutes.
Sex hormones are transported by a plasma protein known as sex-hormone binding
globulin.
The concentration of transcortin increases in pregnancy, but in hepatic cirrhosis,
nephrosis and multiple myeloma decreases.
Bound hormones work as a pool from where free hormones are released. Anytime
the concentration of free hormones decreases that will stimulate the synthesis of hormones
and increases the secretion of corticotropin releasing hormone (CRH).
In liver cortisol is conjugated with glucoronic acid and to a lesser extent to sulphates.
A certain amount of cortisol is transformed in liver into cortison, which is also
gluconoconjugated. They become hydrosoluble and released in blood from where they are
transported to the kidney and excreted through urine. In the urine they are found as 17-
hydroxysteroids. Daily are secreted 2 –12 mg through the urine. Around 10% of cortisol is
transformed in liver into 17-ketosteroid which is also excreted though urine as a
sulfoconjugated product. When the capacity of liver to metabolize or inactivate these
hormones is decreased, these substances may accumulate in blood.
Aldosterone is glucoronoconjugated in liver. Adrenal androgens or sexual hormones
are reverted 17-ketosteroid and they are also excreted into urine.
The normal concentration of aldosterone in blood is about 6 nanograms/dL, and the
average secretory rate is approximately 150 μg/day. The concentration of cortisol in the blood
averages 12 μg/dL, and the secretory rate reaches 15 to 20 mg/day.

Effects of adrenal hormones

Glucocorticoids (GC)- cortisol, corticosterone, cortisone

GC are secreted mainly by zone fasciculate. A small quantity of GC is also secreted by

zona reticularis. Humans secrete about 10 times more cortisol than corticosterone, and
corticosterone has only one fifth of the glucocorticoid activity of cortisol. Cortisol is considered
the physiologically important glucocorticoid in humans. Compared with the glucocorticoids, a

39
much smaller amount of aldosterone is secreted each day. Because of similarities in their
structures, the glucocorticoids and aldosterone have overlapping actions.
Cortisol exerts its effects by interacting with intracellular receptors in target cells.
Because cortisol is lipid soluble, it can easily diffuse through the cell membrane. Once inside
the cell, cortisol binds with its protein receptor in the cytoplasm, and the hormone-receptor
complex then interacts with specific regulatory DNA sequences, called glucocorticoid response
elements, to induce or repress gene transcription. Other proteins in the cell, called
transcription factors, are also necessary for the hormone-receptor complex to interact
appropriately with the glucocorticoid response elements.
Cortisol stimulates gluconeogenesis. It increases the enzymes required to convert
amino acids into glucose in the liver cells and it causes mobilization of amino acids from the
extrahepatic tissues mainly from muscle. Cortisol increases the blood glucose level also by
inhibiting glucose uptake and utilization by peripheral cells.
Studies in isolated tissues have demonstrated that cortisol depresses amino acid
transport into muscle cells and into other extrahepatic cells. The decreased transport of
amino acids into extrahepatic cells lowers their intracellular concentration and consequently
decreases the synthesis of protein. Therefore, cortisol mobilizes amino acids from the
nonhepatic tissues and in doing so diminishes the tissue stores of protein. GC increases the
transport of aa into hepatic cells, where they are used for the synthesis of proteins, plasma
proteins and for gluconeogenesis.
Cortisol reduces the protein stores in essentially all body cells except those of the
liver. This is caused by both decreased protein synthesis and increased catabolism of protein
in the cells. Both these effects may result from decreased amino acid transport into
extrahepatic tissues. Cortisol also depresses the formation of RNA and subsequent protein
synthesis in many extrahepatic tissues, especially in muscle and lymphoid tissue. Together
with the reduced proteins elsewhere in the body, the liver proteins become enhanced.
Cortisol promotes mobilization of fatty acids from adipose tissue. This increases the
concentration of free fatty acids in the plasma, which also increases their utilization for
energy. Cortisol also seems to have a direct effect to enhance the oxidation of fatty acids in
the cells. It has a ketogenic effect.
GC enhace the retention of sodium and to lesser exetent increase the excretion of
potassium. They stimukate the activity of osteoclasts (bone resorption) and inhibit the
osteoblastic activity (bone formation and mineralization). The result of their hypersecretion is
osteoporosis.
It maintains the vascular tonus and increases the force of contraction of heart
because it increases the response of smooth muscle cells to catecholamines. The effect of
catecholamines on heart and smooth muscle cells of vessels is increased by cortisol. GC
maintain plasmatic volume by increasing the flowing of fluid from interstitial space to the
vessels. During neonate period, cortisol inhibits the proliferation of cells in central nervous
system, decreases the mass of brain, the formation of synapses. GC have an inhibitory role on
the morphology and function of nervous system. In adult, the hypersecretion of glucocorticoid
hormones is associated with emotional disturbance that can evoluate until psychosis. The
pattern EEG is changed under the action of cortisol, with predomination of some slow waves.
There is an increase of taste and olfactory sensitivities, tiredness, and incapacity of
concentration.

40
 Glucocorticoids have effect on other glands. They inhibit the secretion of growth
hormone, of TSH, the convertion of tyroxine into T3. They stimulate the effects of glucagon,
the beta adrenergic effect of catecholamines. They inhibit the secretion of vasopressin.
On gastro intestinal tract, glucocorticoid hormones stimulate the secretion of
hydrochloric acid and pepsin. They inhibit the synthesis of prostaglandins, and by these two
effects, they favorise the appearance of peptic ulcer. A side effect of treatment with synthetic
glucocorticoid hormone, Prednison, can produce petic ulcer or gastritis.
On blood cells: they decrease the number of eosinophils, basophils, and lymphocytes
(eosinopenia, basopenia, lymphocytopenia), but they increase the number of neutrophils
(neutrophilia), red blood cells, and of platelets. Because they stimulate the catabolism of
proteins from lymphatic tissue, they decrease the size of lymph nodes, thymus, and they
inhibit the division of lymphocytes. By this they determine lymphocytopenia. They inhibit the
secretion of interleukin 2, produced by helper T lymphocytes. This mediates the
transformation of lymphocytes B into plasmocytes. This explains the immunosuppressive role
of glucocorticoids.
Cortisol decreases the permeability of the capillaries. Cortisol decreases both
migration of white blood cells into the inflamed area and phagocytosis of the damaged cells.
Cortisol suppresses the immune system, causing lymphocyte reproduction to decrease
markedly. Cortisol attenuates fever mainly because it reduces the release of interleukin-1
from the white blood cells. Cortisol increases the production of red blood cells by mechanisms
that are unclear. When excess cortisol is secreted by the adrenal glands, polycythemia often
results, and conversely, when the adrenal glands secrete no cortisol, anemia occcurs.
Anti-inflammatory role is explained by inhibition of phospholipase A, by which is
inhibited the release of arachidonic acid from the phospholipids of cell membrane.
Leukotriens and prostaglandines are pro-inflammatory molecules, produced from arachidonic
acid. Cortisol inhibits the secretion of IL2, of gamma interferon, and the secretion of IL1
produced by macrophages during inflammatory reaction (fever). Glucocorticoid hormones
stabilizes the lysosomal membrane. By inhibition of degranulation of mast cells, histamine,
heparine, serotonin, and other mediators are not released, and by this is explained the
antiallergic effect of glucocorticoids. All these actions explain why glucocorticoids are used in
treatment of inflammatory and immune diseases.
During stress, the secretion of adrenocorticotrope hormone is stimulated.
Corticotrope releasing hormone (CRH) stimulates the release of ACTH from pituitary gland,
that goes to adrenal glands. Due to the effect of cortisol on catecholamines, there is a
vasoconstriction in skin, and a vasodilatation in muscle, so there is a certain distribution of
blood flow, specially oritented toward muscle which is necessary for muscle activity, and by
this, the organism is held to go toward danger. This is named fight or flight reaction. Cortisol
is a life protective hormone because it helps withstand the stress and trauma in life.

Regulation of glucocorticoid hormones secretion

The secretion of glucocorticoids is regulated by adrenocorticotrope hormone (ACTH).

It is a small peptide and it mediates its effect by means of G protein connected with adenylyl
cyclase. Adrenococrticotrope hormone in its turn is released under the action of corticotropin
releasing hormone (CRH) from hypothalamus. The release of adrenocorticotrope hormone
and corticotropin releasing hormone has an oscilating profile. The highest peak is recorded in

41
the morning before the person wakes up, and the lowest value is in the afternoon/evening.
Adrenocorticotrope hormone is released in pulsatory way. The release lasts 10-20 minutes
(the peaks) and after the secretion decreases.
Cortisol has direct negative feedback effects on (1) the hypothalamus to decrease the
formation of CRH and (2) the anterior pituitary gland to decrease the formation of ACTH. Both
of these feedbacks help regulate the plasma concentration of cortisol. Cortisol secretion is
regulated by a negative feedback mechanism by ACTH. Cortisol prolonged treatment or
another synthetic glucocorticoid hormones, will inhibit the secretion of ACTH, and can lead to
atrophy of adrenal gland and of hypophysis.

Mineralocorticoids

They are represented by aldosterone, deoxycorticosterone and corticosterone.

Aldosterone’s mineralocorticoid activity is about 3000 times greater than that of cortisol, but
the plasma concentration of cortisol is nearly 2000 times that of aldosterone. Aldosterone is
synthesized by cells of zona glomerulosa. It is essential for the life. It stimulates the
reabsorption of sodium at the level of distal tubule and collecting tubule, together with
secretion of potassium or hydrogen. Being a steroid, it is liposoluble and it has its receptors at
the level of cytoplasm. The ligant – receptor complex is transported till nucleus where it
activates transcription of specific genes related to the process of sodium and potassium
transport – Na/K-ATP-ase. Because aldosterone mediates its effect by transcription of
different genes, the effects appear after 35-45 minutes.
The reabsorption of sodium takes place also at the level of salivary glands, sweat
glands, and at the level of gastro intestinal mucosa. This is stimulated by aldosterone.
Aldosterone secretion is not under the controle of ACTH. Its secretion is stimulated
by angiotensin II, by increased potassium level, decreased sodium level in blood and
decrease in ECF volume. It can also be stimulated by adrenocorticotrope hormone, but only
in very high doses. Increase potassemia only with with 0.1 mEq/L wil stimulate the secretion
of aldosterone (normal potassemia: 3.5-5 mEq/L). Sodium has less effect.
Deoxycorticosterone is under the control of ACTH.
Angiotensin II is formed by release of renin from juxtaglomerular cells. Angiotensin II
will go to zona glomerulosa. Renin is released from juxtaglomerular cells anytime the blood
flow to kidnbey decreases, blood pressure decreases, when the amount of sodium chlorides
at the level of macula densa decreases. Stimulation of sympathetic nervous system increases
release of renin.
Increase of potassium concentration will depolarize the cells from zona glomerulosa
and by this calcium voltage gated channels are open. Calcium enters the cells and aldosterone
secretion is activated. Aldosterone can have some effects on smooth muscle cells but not
mediated by transcription of genes. This effect is not very clear understood yet.
Aldosterone not only causes potassium to be secreted into the tubules in exchange
for sodium reabsorption in the principal cells of the renal collecting tubules, but also causes
secretion of hydrogen ions in exchange for sodium in the intercalated cells of the cortical
collecting tubules. This decreases the hydrogen ion concentration in the extracellular fluid,
causing a mild degree of alkalosis.
Aldosterone has almost the same effects on sweat glands and salivary glands as it has
on the renal tubules. Both these glands form a primary secretion that contains large quantities

42
of sodium chloride. Much of the sodium chloride, on passing through the excretory ducts, is
reabsorbed, whereas potassium and bicarbonate ions are secreted. Aldosterone greatly
increases the reabsorption of sodium chloride and the secretion of potassium by the ducts.
The effect on the sweat glands is important to conserve body salt in hot environments. The
effect on the salivary glands is necessary to conserve salt when excessive quantities of saliva
are lost.
Aldosterone greatly enhances sodium absorption by the intestines, especially in the
colon, which prevents loss of sodium in the stools. Conversely, in the absence of aldosterone,
sodium absorption can be poor, leading to failure to absorb chloride and other anions and
water as well. The unabsorbed sodium chloride and water then lead to diarrhea, with further
loss of salt from the body.

Adrenal sex hormones (ASH)

ASH secreted by adrenal gland are: dehydroepiandrosteron, androstenedione,

testosterone, estrogen, progesterone. They are secreted by zona reticularis and very little by
zona fasciculata. Their secretion is under the control of adrenocorticotrope hormone and not
under gonadotrope hormones (LH and FSH). In normal conditions ASH have insignificant
physiological effects, due to the low amount of secretion, both in male and female.
Much of the growth of the pubic and axillary hair in the female results from the action
of these hormones. The adrenal cortex is the main source of androgens in the blood in human
females. In the human male, however, androgens produced by the testes and adrenal cortex
contribute to the male sex hormones circulating in the blood. Adrenal androgens normally
have little physiological effect other than a role in development before the start of puberty in
both girls and boys. This is because the male sex hormone activity of the adrenal androgens is
weak. Exceptions occur in individuals who produce inappropriately large amounts of certain
adrenal androgens as a result of diseases affecting the pathways of steroid biosynthesis in the
adrenal cortex.

Disturbances of adrenal gland secretion

Cushing disease or syndrome. In this disease there is a hypersecretion of cortisol. It

is usually due to a tumor secreting ACTH at the level of pituitary gland. It can also be a tumor
at the level of adrenal gland. Because of the high secretion of coritisol, protein metabolism is
increased, especially at the level of muscle, skin and subcutaneous tissue. Because there is an
increase of catabolism, the skin will become thinner, the elastic tissue from skin will be
catabolized and through skin the vascularization can be seeen (verbix). Lipids are mobilized
from the areas where they are stored to other areas. Lipids are stored at the level of face, neck
and superior part of trunk giving a peculiar aspect : buffalo neck. Because amino acids are
mobilized toward liver and transformed into glucose, in Cushing disease there is
hyperglycemia and diabetes mellitus.
Due to the fact that cortisol has mineralocorticoid effects , water and Na are retained
in the organism. This increases blood pressure. Also osteoporosis is noted because of
acceleration of catabolism of bone matrix. Another aspect of Cushing disease is the
consequence of androgen hormones hypersecretion - acne and hirsutism.
Primary hyperaldosteronism or Conn disease = hypersecretion of aldosterone. It is
due to a tumor at the level of zona glomerulosa. Because of that there is a retention of sodium

43
and high losses of potassium. Potassium is lost in higher concentrations that can lead to a
nephropathy named hypokalemic nephropathy. The kidney has a decreased capacity to
concentrate urine and polyuria occurs. Decrease K and increase Na induce alkalosis, followed
by decreases of plasma calcium (spasmophilia and thetahy ).
Secondary hyperaldosteronism is not the result of a tumor, but of other conditions
in which the secretion of aldosterone is stimulated: dehydration, shock, cardiac failure,
edema, toxemia of pregnancy or hepatic cirrhosis (ascitis).
In Addison disease adrenal gland is partially or totally destroyed. Sometimes this is
an autoimmune disease, or it could be the result of tuberculosis, or tumors or bleeding.
Adrenal hormones are not produced. There will be deficiency of mineralocorticoid and
glucocorticoid. Sodium is lost, potassium saved, blood volume decreases, hypoglycemia is
associated. The compensatory mechanism will be secretion of adrenocorticotrope hormone.
One aspect is hyperpigmentation in some areas, specially elbows, areola mammaris, at the
level of scars, oral mucosa and genital mucosa. In a person with Addison’s disease, the output
of glucocorticoids does not increase during stress. Yet whenever different types of trauma,
disease, or other stresses, such as surgical operations, supervene, a person is likely to have an
acute need for excessive amounts of glucocorticoids and often must be given 10 or more times
the normal quantities of glucocorticoids to prevent death. This critical need for extra
glucocorticoids and the associated severe debility in times of stress is called an addisonian
crisis.

Adrenogenital syndrom

An adrenocortical tumor that secretes excessive quantities of androgens causes

intense masculinizing effects throughout the body. If this occurs in a female, she develops
virile characteristics: growth of a beard, a much deeper voice, baldness, masculine distribution
of hair on the body and the pubis, growth of the clitoris to resemble a penis, and deposition
of proteins in the skin and especially in the muscles to give typical masculine characteristics.
In the prepubertal male, a virilizing adrenal tumor causes the same characteristics as
in the female plus rapid development of the male sexual organs. In the adult male, the virilizing
characteristics of adrenogenital syndrome are usually obscured by the normal virilizing
characteristics of the testosterone secreted by the testes. It is often difficult to make a
diagnosis of adrenogenital syndrome in the adult male. In adrenogenital syndrome, the
excretion of 17-ketosteroids (which are derived from androgens) in the urine may be 10 to 15
times normal. This finding can be used in diagnosing the disease.

PHYSIOLOGY OF THE MALE REPRODUCTIVE SYSTEM

44
 Testis has two roles :
1) Gametogenesis -production of spermatozoa
2) Endocrine role: synthesis of sex hormones (testosterone)

The reproductive function of the body starts at the puberty : 12 years for girls (8 –13
years), and 14 years for boys (9 -14 years). Until the age of puberty, gonads remain inactive,
but around puberty they express their function. The activation of gonads happens under the
action of gonadotrope hormones (FSH, LH) produced by pituitary gland and a gonadoliberin
released by hypothalamus (GnRH).
During the initial stage, which begins during fetal life and ends in the first year of life,
GnRH, the gonadotropins (LH, FSH), and gonadal sex hormones are secreted at relatively high
levels. From infancy to puberty, the secretion rates of these hormones are very low and
reproductive function is quiescent. At puberty hormonal secretion rates increase very much.
Finally, reproductive function diminishes later in life, largely because the gonads become less
responsive to the gonadotropins.
In newborns there is a secretion of gonadoliberin in hypothalamus, but this secretion
is stopped at the age of 3-6 months. The synchronous and phasic activity of these neurons
from hypothalamus is resumed at the puberty. At the beginning of puberty, the secretion of
gonadoliberin is increased especially during the periods of sleep, but as long as puberty
progresses, the secretion of gonadoliberin is also done during the daytime.
It is not clear why the activity of these neurons is stopped until puberty, but it is
supposed that at the level of hypothalamus there is a mechanism by which the secretion of
gonadoliberin is inhibited until puberty.

Testis

As the embryo develops after fertilization, there is no differentiation of the sexes for
the first six weeks. In the seventh or eighth week, the genetic males start developing testis in
a region close to the adrenal glands inside the abdomen. The testis starts secreting hormones
that cause the development of the male internal and external genitalia. In genetic females,
the absence of Y chromosomes and lack of male hormones is responsible for the development
of the female internal and external genitalia. Normally, the testis has descended by the
seventh month of development and is positioned in the scrotum at birth .
It has two function: 1) to produce spermatozoa, which is gametogenetic function and
2) to synthetize testosterone which is the endocrine function.

1) Spermatogenesis- starts at the puberty

Spermatogenesis is initiated shortly before puberty, under the influence of the rising
levels of gonadotropins and testosterone, and continues throughout life, with a slight decline
during old age. It takes place at the level of seminiferous tubules (ST).
Each testis has 200 –300 lobules. Each lobule containes 1-4 coiled tubules known as
seminiferous tubules. A testicle contains around 400- 600- 900 semineferous tubules, having
the length between 30 -75 cm, the diameter is 150 – 300 μ. Semineferous tubules are
continued with rete testis, vas efferens, epididymis. At the level of epididymis, spermatozoa
become mobile, and here spermatozoa are stored between two ejaculations. The fluid existing
in seminiferous tubule is the result of secretion of Sertoli cells. Its composition is a little

45
different from plasma, because it contains less proteins and glucose, but it contains more
potassium, aspartic acid, glutamic acid, testosterone and estrogens.
From epididymis, spermatozoa pass into deferens duct/vas deferens. The seminal
vesicles are situated on either side of prostate gland. Secretion of seminal vesicles is named
seminal fluid and is emptied into ampulla of vas deferens. The ampulla is continued as
ejaculatory duct, which passes through prostate to form internal urethra. The secretion of the
seminal vesicles is mucoid and viscous in nature, neutral or slighyly alkaline in reaction. It
contains much fructose (the substrate for glycolysis, which is necessary as a metabolic support
for spermatozoa), but also prostaglandins, folic acid, fibrinogen. It is considered that
prostaglandins from the secretion of seminal vesicles can induce the reverse peristaltic
movements of uterus and oviducts, facilitating the progression of spermatozoa towards ovum.
Fibrinogen is converted into the coagulum as soon as semen is ejaculated. The seminal
vesicles contribute about 60% of the semen volume.
The next element through which spermatozoa pass is prostate. Prostate is made up
of 20 – 30 glands (tubuloalveolar), which open separately into the urethra. At the level of
prostate a milky and alkaline secretion is added, that is rich is phosphates, citratres,
coagulation factors and plasminogen.
The clotting enzymes from prostate fluid convert fibrinogen (from seminal vesicles)
into coagulum. These substances (phosphates, citratres) are useful for neutralization of the
acidic pH of vagina, which is 3.5- 4. Sperm become immobile when pH is < 6. Prostate gland
secretion (0.5 mL) includes fibrinolysin, which is responsible for liquefaction of the coagulated
semen 15 to 30 minutes after ejaculation, releasing sperm.
The vas deferens and the portion of the epididymis closest to it serve as a storage
reservoir for sperm, holding them until sexual arousal leads to ejaculation. Also, in the
epididymis the sperm undergo a further testosterone-dependent maturation process.
Movement of the sperm as far as the epididymis results from the pressure created mainly by
the continuous formation of fluid by the Sertoli cells back in the seminiferous tubules. The
sperm themselves are nonmotile at this time. During passage through the epididymis, there
occurs a hundredfold concentration of the sperm by fluid absorption from the lumen of the
epididymis. Therefore, as the sperm pass from the end of the epididymis into the vas deferens,
they are a densely packed mass whose transport is no longer a result of fluid movement but
is due to peristaltic contractions of the smooth muscle in the epididymis and vas deferens.
Maturation of 1 spermatozoon needs 74 days. The volume of semen ejaculated once
is normally is between 2-4 mL. In every mL of semen, there are between 60-100/150 million
of spermatozoa. When the number of spermatozoa decreases, becoming lower than 20
millions/mL almost all of these people are sterile. Often, the decrease of number is correlated
to an abnormal structure of the spermatozoa. Normaly sperm move in a fluid medium at a
velocity of 1 to 4 mm/min.
The semen contains only 10% sperm by volume, with the remainder consisting of the
combined secretions of the accessory glands (60% from the seminal vesicles and 30% from
prostate).
Number of spermatozoa = 60 –100/150 million/mL
Lower than 20 milliion sperms/mL = oligozoospermia
Lack of sperm in semen = azoospermia

46
 In order to have a normal spermatogenesis, it is necessary for the T° of testis to be
decreased (32°C). The position of testis may be changed by the contraction of cremasterian
muscles, according to the temperature of environment.
Spermatogenesis starts with some immature cells called spermatogonia ( they begin
to divide mitotically at puberty - they have 23 pairs of chromosomes) that pass through
different stages. During the proliferative stage spermatogonia divide by mitosis, without any
change in chromosomial number. In men there are usually seven generations of
spermatogonia. They transform into primary spermatocytes (cells that will undergo the first
meiotic division- they still have 46 pairs of chrm.), secondary spermatocytes (which contains
23 two-chromatid chromosomes, it suffers the second meiotic division), then spermatids, and
finaly from spermatids are formed spermatozoa. Thus, each primary spermatocyte,
containing 46 two-chromatid chromosomes, gives rise to four spermatids, each containing 23
one-chromatid chromosomes.
Spermatozoon/sperm is 60 μ long. It consists of 4 parts: head, neck, body, tail.
The head of a sperm is oval in shape, consists almost entirely of the nucleus, which
contains the DNA bearing the sperm’s genetic information. The tip of the nucleus is covered
by the acrosome, a protein-filled vesicle containing several enzymes that plays an important
role in the sperm’s penetration of the egg (proteolytic enzymes, hyaluronidase, acid
phosphatase, mucoplysaccharides). The head is connected to the body by a short neck, which
is made up of anterior and posterior end knobs. Often the neck and body are called midpiece.
The body is cylindrical (5 - 9 μ). It consists of central core called axial filament, covered by the
cytoplasmic capsule. The axial filament passes through the body and a perforated disc called
end disc or end ring centriole and reaches the tail as axial thread. In the body the axial filament
is surrounded by a spiral filament made up of mitochondria. The tail consists of two pieces: a)
the chief or main piece of tail, long of 40-50 μ; it is enclosed by cytoplasmic capule and has an
axial thread. b) the terminal or end piece of tail - it has only the axial filament. Most of the
tail is a flagellum—a group of contractile filaments that produce whiplike movements capable
of propelling the sperm at a velocity of 1 to 4 mm/min. The sperm’s mitochondria form the
midpiece of the sperm and provide the energy for the sperm’s movement.
The normal human male manufactures approximately 30 millions sperms per day.
After ejaculation the survival time is only 24-48 h at a temp. equivalent to body temp.
Inside of ST there are Sertoli cells, which are supporting cells. They are tall and large,
irregular columnar cells extending from BM to lumen of ST. The spermatogenetic cells present
in ST are attached to Sertoli cells by means of cytoplasmic connections.
Sertoli cells contain glycogen, which is necessary for maturation of spermatozoa.
Outside of seminiferous tubules there are other cells - Leydig cells. Sertoli cells have receptors
for FSH and testosteron.

47
Spermatogenesis (Vander et al, Human Physiology: The Mechanism of Body function, eighth
edition, The McGraw Hill Companies, 2001)

There is a barrier between blood and seminiferous tubules (blood-testis barrier). It is

formed by tight junctions between the adjacent Sertoli cells near the basement membrane of
seminiferous tubule. Due to this barrier, the penetration of different proteins and
macromolecules from interstitial space into seminiferous tubules is impeded. However blood-
testes barrier permits some subst. to pass: water, urea, nutritive subst. nec for developing of
spermatozoa, hormones nec. for spermatogenesis. Because of this spermatozoa are outside
the immune system. This barrier is organized beginning with puberty. Because spermatozoa
do not come in contact with immune system, spermatozoa work like some antigens for the
immune system. It is described a type of infertility in which antibodies against the own sperm
are formed.
Sertoli cells synthesize several proteins: androgen binding protein (testosterone
binding protein) by which testosterone is bound. Sertoli cells also convert androgens into
estrogen (aromatase), secrete estrogen binding protein (EBP), inhibin (it inhibits the release
of FSH from hypophysis), activin, Mullerian regression factor (MRF) (it is responsible for the
regression of Mullerian duct during sex differentiation in fetus).
Sertoli cells phagocytose residual bodies (excess cytoplasm resulting from the
transformation of spermatids to spermatozoa) and damaged germ cells, provide structural
support and nutrition for germ cells, secrete fluid, and assist in spermiation, the final
detachment of mature spermatozoa from the Sertoli cell into the lumen. Spermiation may
involve plasminogen activator, which converts plasminogen to plasmin, a proteolytic enzyme
that assists in the release of the mature sperm into the lumen. Sertoli cells also synthesize
large amounts of transferrin, an iron-transport protein important for sperm development.
Androgen-binding protein (ABP) is a 90-kDa protein, made of a heavy and a light chain,
that has a high binding affinity for dihydrotestosterone and testosterone. It is similar in
function with sex hormone-binding globulin (SHBG), synthesized in the liver. ABP is found at

48
high concentrations in the human testes and epididymis. It serves as a carrier of testosterone
in Sertoli cells, as a storage protein for androgens in the seminiferous tubules and as a carrier
of testosterone from the testis to the epididymis.
Leydig cells are interstitial cells that lie in the small connective tissue space between
the tubules. They do not have FSH receptors, but FSH can increase the number of developing
Leydig cells by stimulating the production of growth stimulators from Sertoli cells that
subsequently enhance the growth of the Leydig cells. In addition, androgens stimulate the
proliferation of developing Leydig cells. Estrogen receptors are present on Leydig cells, and
they reduce the proliferation and activity of these cells.
Leydig cells have LH receptors. The major effect of LH is to stimulate androgen
secretion via a cAMP-dependent mechanism. The main product of Leydig cells is testosterone,
but two other androgens of less biological activity, dehydrotestosterone and
androstenedione, are also produced.
There are bidirectional interactions between Sertoli and Leydig cells. The Sertoli cells
do not produce testosterone but contain testosterone receptors as well as FSH-dependent
aromatase. The Leydig cell does not produce estradiol, but contains receptors for it, and
estradiol can suppress the response of the Leydig cell to LH. Testosterone diffuses from the
Leydig cells, crosses the basement membrane, enters the Sertoli cell, and binds to ABP. As a
result, androgen levels can reach high local concentrations in the seminiferous tubules.
Testosterone is obligatory for spermatogenesis and the proper functioning of Sertoli cells.
In Sertoli cells, testosterone also serves as a precursor for estradiol production.

Role of hormones in spermatogenesis

Spermatogenesis is influenced by many hormones, that act directly or indirectly.

These are :
- FSH
- Testosterone
- Estrogen
- LH
- GH
- Inhibin
- Activin

FSH stimulates the production of androgen-binding protein and plasminogen activator,

increases secretion of inhibin, and induces aromatase activity for the conversion of androgens
to estrogens. The testosterone receptor is within the nucleus of the Sertoli cell. FSH is
responsible for the initiation of spermatogenesis, by inducing the proliferation of
spermatogonia.
Testosteron – is responsible for the stages of growth, maturation and transformation
of spermatogonia, also for the maintenance of spermatogenesis.
Estrogens - plasma estrogen arises in males from secretion of small amounts by the
testes and from conversion of androgens to estrogen in many nongonadal tissues (notably
adipose tissue). Conversely, in females androgens are secreted, in small amounts, by the
ovaries and, in larger amounts, by the adrenal cortex. (Some of these androgens are then
converted to estrogen in nongonadal tissues, just as in men, and contribute to the plasma
estrogen.)

49
 Estrogen is formed from testosteron in Sertoli cells. It is necessary for the stage of
transformation of spermatides into sperm, together with testosteron.
LH- it is essential for the secretion of testosteron from Leydig cells.
Growth Hormone- necessary for the metabolic processes in testis and for proliferation
of spermatogonia. In pituitary dwarfs, the spermatogenesis is severely affected.
Inhibin – is secreted by Sertoli cells and by granulose cells of ovarian follicles. Its secr.
is stimulated by FSH. Inhibin inhibits FSH secretion through feedback mecs. FSH induces
spermatogenesis by stimulating Sertoli cells. So, when the rate of spermatogenesis increases,
there is simultaneous increase in inhibin secretion, too. Inhibin in turn acts on anterior
pituitary and inhibits secretion of FSH leading to decrease in the pace of spermatogenesis. It
is believed that inhibin also inhibits FSH secretion indirectly by inhibiting GnRH secretion from
hypothalamus.
Activin – the exact location of its secretion in testis is not known (might be Sertoli and
Leydig cells). It has opposite actions to inhibin. It increases secr. of FSH and accelerates
spermatogenesis.
Spermatogenesis requires high intratesticular levels of testosterone, secreted from the
LH-stimulated Leydig cells. The testosterone diffuses across the basement membrane of the
seminiferous tubule, crosses the blood-testis barrier, and complexes with ABP. Sertoli cells,
but not spermatogenic cells, contain receptors for testosterone. Sertoli cells also contain FSH
receptors. However, recent studies using mice, in which the alpha subunit of FSH has been
mutated to an inactive form, reveal that the testes are small but do produce sperm. The
absolute requirement for FSH in sperm production remains unknown. From these data, it
appears that testosterone may be sufficient for spermatogenesis.
Upon entering meiosis, spermatogenesis appears to depend on the availability of FSH
and testosterone. In human males, FSH is thought to be required for the initiation of
spermatogenesis before puberty. When adequate sperm production has been achieved, LH
alone (through stimulation of testosterone production) or testosterone alone is sufficient to
maintain spermatogenesis.

2) Testicular steroidogenesis - endocrine role

Testis secretes male sex hormones, called androgens: testosteron (secreted in large
quantities), dihydrotestosterone and androstenedione. Activin and inhibin are also secreted,
but they do not have androgenic effects.

Testosteron is a C19 steroid, synthesized and seed by the interstitial cells of Leydig, which
lie in the interstices between the seminiferous tubules and constitute about 20% the mass of
the adult testes. Leydig cells are almost nonexistent in the testes during childhood when the
testes secrete almost no testosterone, but they are numerous in the newborn male infant for
the first few months of life and in the adult male any time after puberty; at both these times
the testes secrete large quantities of testosterone.
The action of LH on Leydig cells is mediated through specific LH receptors on the plasma
membrane. A Leydig cell has about 15,000 LH receptors, and occupancy of less than 5% of
these is sufficient for maximal steroidogenesis. LH stimulates steroidogenesis by two principal
activations. One is the phosphorylation of cholesterol esterase, which releases cholesterol
from its intracellular stores. The other is the activation of CYP11A1 (cholesterol side-chain
cleavage enzyme). Leydig cells also contain receptors for prolactin (PRL). Hyperprolactinemia
50
in men with pituitary tumors, usually microadenomas, is associated with decreased
testosterone levels. This condition is due to a direct effect of elevated circulating levels of PRL
on Leydig cells, reducing the number of LH receptors or inhibiting downstream signaling
events. In addition, hyperprolactinemia may decrease LH secretion by reducing the pulsatile
nature of its release. Under nonpathological conditions, however, PRL may synergize with LH
to stimulate testosterone production by increasing the number of LH receptors.

Plasma level of testosteron (T)

An adult man produces 6 to 7 mg testosterone per day. This amount slowly declines
after age 50 and reaches about 4 mg/day in the seventh decade of life. (T) in male is 3 - 10
ng/ml, in female is < 1 ng/ml.
After secretion by the testes, about 97% of the testosterone becomes either loosely
bound with plasma albumin (1/3) or more tightly bound with a beta globulin called sex
hormone–binding globulin (2/3) and circulates in the blood in these states for 30 minutes to
several hours. SHBG binds both estradiol and testosterone, with a higher binding affinity for
testosterone.
In many target tissues, T is converted into dehydrotestosterone which is the most
active androgen. In adipouse tissue, liver and hypothalamus T is converted into estradiol. T is
matebolized in liver - it is converted into inactive forms of androsterone and
dehydroepiandrosterone and simultaneously conjugated with either glucuronides or sulfates.
These are excreted either into the intestin by way of bile or into the urine through the kidneys.

Regulation of testosteron secretion

Hypothalamic neurons produce gonadotropin-releasing hormone (GnRH), which

regulates the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH).
The neurons that produce GnRH are found in the infundibulum and arcuate nucleus. GnRH
enters the hypothalamic-pituitary portal system and binds to receptors on the plasma
membranes of pituitary cells, resulting in the synthesis and release of LH and FSH. LH and FSH
bind membrane receptors on Leydig and Sertoli cells. The activation of LH and FSH receptors
on these cells increases the intracellular second messenger cAMP. cAMP binds to protein
kinase A, which activates transcription factors such as steroidogenic factor-1 (SF-1) and cAMP
response element binding protein (CREB). These factors activate the promoter region of the
genes of steroidogenic enzymes that control testosterone production by Leydig cells. Similar
signal-transducing events occur in Sertoli cells that regulate the production of estradiol. The
testis converts testosterone and some other androgens to estradiol by the process of
aromatization, although estradiol production is low in males.
GnRH is secreted intermittently a few minutes at a time once every 1 to 3 hours. GnRH
pulsatility is ultimately necessary for proper functioning of the testes because it regulates the
secretion of FSH and LH, which are also released in a pulsatile fashion. Continuous exposure
of gonadotrophs to GnRH results in desensitization of GnRH receptors, leading to a decrease
in LH and FSH release.
FSH binds with specific FSH receptors attached to the Sertoli cells in the seminiferous
tubules. This causes these cells to grow and secrete various spermatogenic substances.
Simultaneously, testosterone (and dihydrotestosterone) diffuses into the seminiferous
tubules from the Leydig cells and performs a strong tropic effect on spermatogenesis.

51
 Testosterone, estradiol, inhibin, activin, and follistatin are major testicular hormones
that regulate the release of the gonadotropins LH and FSH. Generally, testosterone, estradiol,
and inhibin reduce the secretion of LH and FSH in the male. Activin stimulates the secretion of
FSH, whereas follistatin inhibits FSH secretion. Testosterone inhibits LH release by decreasing
the secretion of GnRH and, to a lesser extent, by reducing gonadotroph sensitivity to GnRH.
Estradiol formed from testosterone by aromatase also has an inhibitory effect on GnRH
secretion. Acute testosterone treatment does not alter pituitary responsiveness to GnRH, but
prolonged exposure significantly reduces the secretory response to GnRH.
Inhibin is produced by Sertoli cells. It acts directly on the anterior pituitary and inhibits
the secretion of FSH but not LH.
Activin is produced by Sertoli cells, it stimulates the secretion of FSH.
Follistatin is a 31 to 45 kDa single-chain protein hormone, with several isoforms, that
binds and deactivates activin. Thus, the deactivation of activin by binding to follistatin reduces
FSH secretion. Follistatin is apparently produced by Sertoli cells and acts as a paracrine factor
on the developing spermatogenic cells.

Effects of testosteron

T is responsible for the distinguishing characters of masculine body. In the fetal life, the
testes are stimulated by human chorionic gonadotroprin secreted by placenta. In childhood,
practically no T is secreted until the age of 10-12 y. Afterwards, the secretion of T starts, it
increases rapidly at the onset of puberty and lasts through most of the remaining part of life.
The secretion decreases after 40 years.

Role of T in fetal life

T is secreted durinf fetal life beginning with the 2-4 month. It is involrved in : a) sex
differentiation, b) development of accessory sex organs, c) descent of testes.

a) Sex chromosomes are responsible for determination of the sex of the person and
T for sex differentiation of fetus. In fetus there are two ducts : Mullerian duct (
gives rise to female accessory sex organs) and Wolffian duct ( gives rise to male
accessory sex oragns) . If T is secreted by the genital ridge of fetus at 7 th week
of pregnancy, the mullerian duct disappears and male sex organs develop.

b) Development of accessory sex organs and external genitalia - T is imp. for growth
of penis , scrotum, genital ducts, seminal vesicle and prostate.

c) Descent of testes - from abdominal cavity into scrotum through the inginal canal
just before birth.

Role of T in adult life

1. Required for initiation and maintenance of spermatogenesis (acts via Sertoli cells)
2. Decreases GnRH secretion via an action on the hypothalamus
3. Inhibits LH secretion via an action on the anterior pituitary

52
4. Induces differentiation of male accessory reproductive organs and maintains their
function
5. Induces male secondary sex characteristics; opposes action of estrogen on breast growth
6. Stimulates protein anabolism, bone growth, and cessation of bone growth
7. Required for sex drive and may enhance aggressive behavior
8. Stimulates erythropoietin secretion by the kidneys

 On sex organs - T increaseas the size of penis , scrotum and testes at puberty . Since
puberty to 20 y these organs increase of 8 times. The fetal differentiation, and later growth
and function of the entire male duct system, glands, and penis all depend upon testosterone.
Virtually all the male secondary sex characteristics are dependent on testosterone. T
stimulates the growth and secretory activity of the epididymis and accessory glands, and
increases the pigmentation of the genitalia. Enlargement of the testes occurs under the
influence of the gonadotropins (LH and FSH). Throughout adulthood, androgens are
responsible for maintaining the structural and functional integrity of all reproductive tissues.
Castration of adult men results in regression of the reproductive tract and involution of the
accessory glands.

 In other target cells (brain) testosterone is transformed not to dihydrotestosterone

but to estradiol, which is the active hormone in these cells. Depending on the target cells,
testosterone acts as testosterone per se, dihydrotestosterone, or estradiol. This has important
pathophysiological implications since some men lack the enzymes required to perform one or
the other of the transformations of testosterone. Therefore, they will exhibit certain signs of
testosterone deficiency but not others. For example, a man with absence of the enzyme for
forming dihydrotestosterone will have normal differentiation of his reproductive duct
structures (an effect of testosterone per se) but lack of development of his external genitalia
(an effect requiring dihydrotestosterone).
 Secondary Sex Characteristics and Growth - other testosterone dependent secondary
sexual characteristics are: deepening of the voice resulting from the growth of the larynx, thick
secretion of the skin oil glands (this predisposes to acne), and the masculine pattern of fat
distribution. Testosterone also stimulates bone growth, indirectly through its stimulation of
growth hormone secretion, but ultimately shuts off bone growth by causing closure of the
bones’ epiphyseal plates. Testosterone is an “anabolic steroid” in that it exerts a direct
stimulatory effect on protein synthesis in muscle. T accelerates the transport of amino acids
in muscle, synthesis and storage of proteins in muscle and decreases breakdown of proteins.
After puberty T incr. the thickness of bones with deposition of Ca. It increases the bone
matrix because of the anabolic role of it on protein . If testes are removed before puberty -->
delayed fusion  height of the person increases.
T incr. the thickness of skin over the entire body, the ruggedness of subcutaneous tissue,
due to the deposition of proteins in skin.
Hair is classified by its sensitivity to androgens into nonsexual (eyebrows and
extremities); ambisexual (axilla), which is responsive to low levels of androgens; and sexual
(face, chest, upper pubic triangle), which is responsive only to high androgen levels. Hair
follicles metabolize testosterone to DHT or androstenedione. Androgens stimulate the growth
of facial, chest, and axillary hair; however, along with genetic factors, they also promote
temporal hair recession and loss. Normal axillary and pubic hair growth in women is also under

53
androgenic control, whereas excess androgen production in women causes the excessive
growth of sexual hair (hirsutism).
T increases BMR to 10- 15% due to anabolic effect on prot. It incr. Na reabs. from renal
tubule . Testosterone is necessary for expression of the genetic determinant of the common
type of baldness (“male pattern”) in men. Testosterone stimulates the secretion of the
hormone erythropoietin by the kidneys, and this is a major reason that men have a higher
hematocrit than women.
 Behavior - Testosterone is essential in males for development of sex drive at
puberty. It also plays an important role in maintaining sex drive in the adult male.
Many sites in the brain contain androgen receptors, with the highest density in the
hypothalamus, preoptic area, septum, and amygdala. Most of those areas also contain
aromatase and many of the androgenic actions in the brain result from the aromatization of
androgens to estrogens. The pituitary also has abundant androgen receptors, but no
aromatase.

Erection

Erection is a vascular phenomenon, during which the penis becomes rigid. The penis
consists of three cylindrical vascular compartments running its entire length. Normally the
small arteries supplying the vascular compartments are constricted so that the compartments
contain little blood and the penis is flaccid. During sexual excitation, the small arteries dilate,
the three vascular compartments become engorged with blood at high pressure, and the penis
becomes rigid.
The vascular dilation is initiated by neural input to the small arteries of the penis.
Moreover, the veins emptying them are passively compressed, thus contributing to the
engorgement. This entire process occurs rapidly, complete erection sometimes taking only 5
to 10 s. At rest, the dominant input is via sympathetic neurons; they release norepinephrine,
which causes the arterial smooth muscle to contract. During erection the sympathetic input
is inhibited, but much more important is the activation of nonadrenergic, noncholinergic
autonomic neurons to the arteries (NO) . These neurons release nitric oxide, which relaxes
the arterial smooth muscle.
The primary stimulus comes from mechanoreceptors in the genital region, particularly
in the head of the penis. The afferent fibers (internal pudendal nerve) carrying the impulses
synapse in the lower spinal cord on interneurons that control the efferent outflow (pelvic
splanhnic nerve- nervi erigenetes).
Higher brain centers, via descending pathways, may also exert profound stimulatory or
inhibitory effects upon the autonomic neurons to the small arteries of the penis. Thus,
mechanical stimuli from areas other than the penis, as well as thoughts, emotions, sight, and
odors, can induce erection in the complete absence of penile stimulation (or prevent erection
even though stimulation is present).

Ejaculation

The discharge of semen from the penis—ejaculation— is also a spinal reflex, the
afferent pathways from penile mechanoreceptors being identical to those for erection. When
the level of stimulation is high enough, there is elicited a patterned sequence of discharge of
the efferent neurons. This sequence can be divided into two phases: (1) The smooth muscles

54
of the epididymis, vas deferens, ejaculatory ducts, prostate, and seminal vesicles contract as
a result of sympathetic stimulation, emptying the sperm and glandular secretions into the
urethra (emission); and (2) the semen is then expelled from the urethra by a series of rapid
contractions of the urethral smooth muscle as well as the skeletal muscle at the base of the
penis. During ejaculation, the sphincter at the base of the urinary bladder is closed so that
sperm cannot enter the bladder nor can urine be expelled from it.
The rhythmical muscular contractions that occur during ejaculation are associated with
intense pleasure and many systemic physiological changes (increase the heart rate and blood
pressure), the entire event being termed as orgasm.

Disturbances of the male reproductive function

Cryptorchidism – is a congenital disorder in which one or both testes do not descend

from abdomen into scrotum. Males with untreated testes are prone for testicular cancer.
Extirpation of testes before puberty- eunuchism (the sex organs do not increase in size, the
secondary sexual characteristiscs do not develop, the persons are taller, there is an abnormal
deposition of fat on buttock, hips, breast and pubis). Extirpation of testes in adults- does not
cause loss of secondary sex chracteristics, but accessory sex organs start degenerating.
Erection occurs but ejaculatgion is rare due to degeneration of accessory sex organs and lack
of sperms. Hypogonadism in males – reduction of the activity of gonads (due to
cryptorchidism, castration, under developed testes, disorders of gonadotrops,
hypothalamus). The symptoms are similar to those developed after extirpation of testes.
Hypergonadism – hypersecr. of sex hormones by gonads, usually due to the tumor of Leydig
cells. It is common in prepubertal boys who develop precocious pseudopuberty.

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