REPRODUCTIVE HORMONES

Larry J. Ream, Ph.D.

Department of Neuroscience, Cell Biology and Physiology
105A White Hall
775-3188
larry.ream@wright.edu
READING ASSIGNMENT
Preview Chapters 10, 17 and 18 in Netters Essential Histology.
OBJECTIVES

List the different types of gonadotropin hormones and their respective functions.
List the different glands/tissues that produce gonadotropin hormones.
Describe the endocrine regulation of testicular and ovarian function by GnRH, FSH,
LH, and inhibin.
List the different types of steroid hormones and their respective functions.
Identify the structure of steroid hormones.
Explain the five stages of synthesis of steroid hormones.
Describe the regulation of steroid hormones.
Describe the cellular mechanisms of action of steroid hormones.

PROTEIN SEX HORMONES

GONADOTROPHIC HORMONES
LH and FSH are glycoproteins that regulate development, growth, pubertal maturation,
reproductive processes, and sex steroid hormone secretion of the gonads of either sex.
Both hormones are usually secreted by a single cell type, the gonadotroph (a type of
basophil). Gonadotrophs make up about 15% of the anterior pituitary cell population,
and they are scattered throughout the gland. Small subclasses of gonadotrophs secrete
only LH or only FSH. Gonadotrophs also appear to go through cycles in which
differential expression of the two gonadotropins occurs at different times. Both
hormones are present by 10 or 12 weeks of fetal life; however, neither is absolutely
required for the initial intrauterine development of the gonads or for the initial steps in
sexual differentiation.

 Each hormone is composed of the common pituitary hormone subunit and of a

different subunit that is responsible for the unique biologic activity of each hormone.

FOLLICLE STIMULATING HORMONE

FSH is a glycoprotein with a molecular weight of 33,000.
Each monomeric unit of FSH is a protein molecule with a sugar attached to it; two of
these make the full, functional protein. The structure of FSH is similar to LH, TSH, and
hCG. The protein dimer contains 2 different polypeptide units, and
subunits connected by two disulfide bridges. The subunits of LH, FSH,
TSH, and hCG are identical, and contain 92 amino acids. The subunits
vary. FSH has a beta subunit of 118 amino acids, which confers its specific
biologic action and is responsible for interaction with the FSH-receptor.
The sugar part of the hormone is composed of fucose, galactose, mannose,
galactosamine, glucosamine, and sialic acid, the latter being critical for its biologic halflife (which is 3-4 hours).
Main functions of FSH:
- promote and sustain ovarian follicular growth in women and spermatogenesis
in men
- stimulates the synthesis of its own receptor on the ovarian granulosa cells and
Sertoli cells of the testis, and the LH receptor on granulosa cells
- stimulates aromatase activity inside the granulosa cells (the enzyme
converting androgens into estrogens)
FSH synthesis and secretion by the pituitary controlled by:
- GnRH
- ovarian estrogens
- activin and inhibin
LUTEINIZING HORMONE
LH is a glycoprotein with a molecular weight of 28,000. LH has a subunit of 121
amino acids that confers its specific biologic action and is responsible for interaction with
the LH receptor. This subunit contains the same amino acids in sequence as the

subunit of hCG and both stimulate the same receptor, however, the hCG subunit
contains an additional 24 amino acids, and both hormones differ in the composition of
their sugar moieties. The different composition of these oligosaccharides affects
bioactivity and speed of degradation. The biologic half-life of LH is 20 minutes, shorter
than that of FSH (3-4 hours) or hCG (24 hours).
Main functions of LH:
- promoting androgen synthesis in the ovarian thecal cells and in the interstitial
cells of the testes
- induces ovulation by stimulating the cascade of proteolytic enzymes leading
the rupture of the mature Graafian follicle
- maintains the corpus luteum during the menstrual cycle
LH synthesis and secretion by the pituitary is controlled by several factors:
- GnRH
- ovarian estrogens and progesterone
SECRETION OF LH AND FSH
The regulation of FSH and LH embodies pulsatile, periodic, diurnal, cyclic, and stageof-life elements. Regulation is also different in women and men.
The secretions of both LH and FS are stimulated primarily by a single hypothalamic
hormone, gonadotropin-releasing hormone. The cells of origin of GnRH are
predominantly in the arcuate nucleus and the preoptic area of the hypothalamus.
Most projections of the GnRH neurons terminate in the median eminence. The GnRH
released from these terminals is picked up by the capillaries of the portal vessels and
transported to the anterior pituitary.
GnRH is a decapeptide that is initially synthesized as a large precursor prohormone that
also yields other products. The preproGnRH consists of a 23-amino acid signal peptide, a
10-amino acid GnRH, and a 56-amino acid GnRH-associated peptide (GAP). The GnRH
is flanked by pairs of basic amino acids that designate enzymatic cleavage sites.

 The general framework for the hypothalamic-pituitary-gonadal axis is presented in the

figure below.

Other protein
products of
the gonads
influence
FSH secretion

Various influences regulate the release of GnRH. Negative modulation by endorphins

and dopamine may mediate some of the steroid hormone feedback.
GnRH causes a much greater increase in LH than in FSH secretion.
Pulsatile secretion of LH, which is due mainly to pulsatile secretion of GnRH, does not
depend on the presence of sex steroid hormones from target glands (see below left).

 FSH secretion also exhibits a pulsatile pattern that is generally synchronized with the
pattern of LH secretion but is of lesser magnitude (see above right).
The secretion of LH and FSH is regulated by testosterone and estradiol; the basic
regulatory mechanism is classic negative feedback.
Other protein products of the gonads influence FSH secretion. Separate gonadal protein
products selectively suppress (inhibin and follistatin) or stimulate (activin) FSH release.
These protein products may be expressed locally in the pituitary gland and exert their
effects by paracrine and autocrine actions.
Binding of GnRH to membrane receptors increases intracellular Ca++ levels and
phosphorylation of proteins, which together lead to increased LH release. The rise in
intracellular Ca++ occurs via two mechanisms (see below): stimulation of release of Ca++
from intracellular storage sites and increased Ca++ influx from the extracellular fluid.

INHIBIN, ACTIVIN AND FOLLISTATIN

In addition to the traditional feedback inhibition of gonadotropin hormone release, locally
produced factors are also involved in regulation of gonadotropin release.
Inhibin is produced and released by Sertoli cells in response to FSH stimulation and
exerts both paracrine and endocrine responses. Inhibin belongs to the transforming
growth factor- (TGF-) superfamily of growth factors. Inhibins are heterodimer
glycoproteins consisting of an - and -subunit (A or B). Of the two forms of inhibin
(-A and -B), inhibin B is the physiologically important form in males. Its main
function is to suppress the secretion of FSH from the pituitary in a classic negative
feedback endocrine mechanism through binding to a membrane-spanning
serine/threonine kinase receptor. Inhibin B secretion appears to be dependent on Sertoli
cell proliferation, maintenance, and spermatogenesis, all of these functions regulated by
FSH. Inhibin B levels correlate with total sperm count and testicular volume and can be
used as an index of spermatogenesis.
Activins, members of the same family of peptides as the inhibins, are homodimers or
heterodimers of the -subunit of the inhibins. They are synthesized in many adult tissues
and cell types, and their receptors have been identified in those same tissues, a pattern
more consistent with autocrine or paracrine mechanisms of action. At the pituitary,
locally produced activin stimulates the synthesis of FSH.
!
Follistatin is a single chain autocrine glycoprotein initially isolated from follicular fluid
and identified as a protein fraction that inhibited FSH secretion from the anterior
pituitary, and so was known as FSH-suppressing protein (FSP). Since then its primary
function has been determined to be the inhibition of activin.

GONADOTROPIN ACTIONS IN THE GONADS

LH and FSH are the coordinate pituitary regulators of gonadal function. Through
negative feedback, their synthesis and secretion are increased by decreases in gonadal
steroids.
The term gonadotropin implies a trophic (i.e., a growth-promoting) effect of FSH
and LH on their target cells. In this regard, both gonadotrophins share sequence
homology and three-dimensional structural features with a variety of growth factors, such
as platelet-derived growth factor and several bone morphogenetic proteins. A cysteine
knot motif, consisting of a cluster of three cysteine bridges, is a characteristic feature of
this ligand family.
LH acts primarily by means of a stimulatory G-protein, adenylate cyclase, and cAMP as
a second messenger. At very high LH concentrations, phosphatidylinositol second
messengers, Ca++, and MAP kinase also transduce LH actions.

 FSH acts on ovarian granulosa cells and testicular Sertoli cells by binding to a PM
receptor. The FSH receptor shares partial homology with the LH receptor. The increase
in cAMP concentration that follows FSH-receptor binding increases the transcription of
the aromatase gene and markedly stimulates estrogen synthesis. FSH also stimulates
synthesis of inhibin and numerous other protein products of Sertoli and granulosa cells.
Another important effect of FSH is to increase the number of LH receptors in granulosa
cells, thereby amplifying their sensitivity to LH.
In addition to their actions on steroidogenesis, LH and FSH produce diverse metabolic
effects on their target gonadal cells. Glucose oxidation and lactic acid production are
increased, effects that may lead to local vasodilation. The long-term tropic effects of the
two hormones entail stimulation of amino acid transport, RNA synthesis, and general
protein synthesis.
AGE-RELATED CHANGES IN GONADOTROPIN SECRETION
The hypothalamic-pituitary-gonadal axis is unique in that it changes throughout the
human life span. Although the patterns of change in females and males differ, there are
certain common aspects.

Note transient peaks during gestation and early infancy and low levels thereafter in
childhood. Women subsequently develop monthly cyclic bursts, with LH exceeding
FSH; men do not. Both genders show increased gonadotropin production after age 50
years, with FSH exceeding LH.

HUMAN CHORIONIC GONADOTROPIN

hCG is a glycoprotein composed of 244 amino acids with a molecular mass of 36,700.
hCG is heterodimeric, with an subunit (92 amino acids) identical to that of LH, FSH,
TSH, and a subunit that is unique to hCG. The -subunit of hCG contains 145 amino
acids, and is encoded by six highly-homologous genes that are arranged in tandem and
inverted pairs on chromosome 19q13.3. The two subunits create a small hydrophobic
core surrounded by a high surface area-to-volume ratio: 2.8 times that of a sphere. The
vast majority of the outer amino acids are hydrophilic.
hCG is produced by the syncytiotrophoblast of the placenta and released into the fetal
and maternal circulation. hCG is the most important peptide hormone produced by the
placenta because it rescues the corpus luteum from degeneration and allows continued
progesterone secretion to support the early pregnancy. It is known as the hormone of
pregnancy and is the basis of the pregnancy test. hCG is detected in serum at day 6-8
after implantation, and its levels peak at 60-90 days of gestation, declining thereafter.
hCG has structural and functional similarity to LH, has a much longer half-life (24 hrs),
and exerts its physiologic effects through binding to the LH receptors.
Due to its highly negative charge, hCG may repel the immune cells of the mother,
protecting the fetus during the first trimester. It has also been hypothesized that hCG
may be a placental link for the development of local maternal immunotolerance. For
example, hCG-treated endometrial cells induce an increase in T cell apoptosis. These
results suggest that hCG may be a link in the development of peritrophoblastic immune
tolerance, and may facilitate the trophoblast invasion, which is known to expedite fetal
development in the endometrium. It has also been suggested that hCG levels are linked
to the severity of morning sickness in pregnant women.
Regulation of hCG release from the placenta is not completely understood, but evidence
indicates that its paracrine regulation involves placenta-derived GnRH, activin, and
inhibin.

HUMAN PLACENTAL LACTOGEN AND GROWTH HORMONE

In addition to hCG, the other major placental peptide, which both are only secreted
during pregnancy, is human chorionic somatomammotropin (hCS).
Levels of hCS (also called human placental lactogen, hPL) are high during
pregnancy. hCS is structurally similar to GH and prolactin.
hCS (hPL) consists of 190 amino acids that are linked by two disulfide bonds. Its
molecular weight is 22,125. Like human growth hormone, HPL is encoded by genes on
chromosome 17q22-24. Its biologic half-life is 15 minutes. The hGH and hPL gene
family is important in the regulation of maternal and fetal metabolism and the growth and
development of the fetus.
hCS is produced by the syncytiotrophoblast and is secreted into both the maternal and
fetal circulations after the sixth week of pregnancy. In the fetus, hCS acts via lactogenic
receptors to modulate embryonic development, regulate intermediary metabolism, and
stimulate the production of IGFs, insulin, adrenocortical hormones, and pulmonary
surfactant. Fatty acids and ketones are important energy sources in the fetus and
placenta, and hCS stimulates production of these substrates.
During pregnancy, hGH-V, a GH variant expressed by the placenta, becomes the
predominant GH in the mother. Starting from the 15th to the 20th week of gestation up to
term of pregnancy, placental GH gradually replaces maternal pituitary GH, which
becomes undetectable. hGH-V stimulates IGF-I production and modulates maternal
intermediary metabolism, increasing the availability of glucose and amino acids to the
fetus.
CORTICOTROPIN-RELEASING HORMONE
CRH is produced by the syncytiotrophoblast and trophoblast cells of the placenta.
CRH is a 41-amino acid peptide derived from a 191-amino acid preprohormone.
The CRH concentration increases exponentially throughout pregnancy and peaks during
labor. Placental production of CRH has been linked to the length of gestation and the
timing of parturition and delivery. CRH is secreted into the maternal circulation in large
amounts during the third trimester of pregnancy and may play an important role in the
onset of labor.

PROLACTIN
Prolactin is a polypeptide hormone synthesized and secreted by lactotrophs (aka
mammotrophs, a type of acidophil) in the anterior pituitary. Lactotrophs account for 1520% of the cell population in the anterior pituitary. However, the percentage increases
dramatically in response to elevated estrogen levels, particularly during pregnancy.
Prolactin is the major hormone responsible for
lactogenesis (milk production) and also participates, along
with estrogen, in breast development. In nonpregnant,
nonlactating females and in males, blood levels of prolactin
are low. However, during pregnancy and lactation, blood
levels of prolactin increase, consistent with the hormones
role in breast development and lactogenesis. Chemically,
prolactin is related to growth hormone, having 198 amino
acids in a single-chain polypeptide with 3 internal disulfide
bridges. Molecular weight is 23,000. Its gene and structure
are homologous to those of hGH, but the molecule has a
major midportion loop.
Synthesis of prolactin proceeds as a prohormone. The N-terminal signal peptide is
cleaved, and transient N-glycosylation takes place before the compound arrives at the
Golgi apparatus. There, the hormone molecules that are subsequently destined to be
stored in granules and released by acute stimuli (or secreted by pregnancy) are
deglycosylated. However, some of the N-glycosylated molecules escape complete
processing, and they are secreted constitutively. These molecules form a major part of
the circulating prolactin in nonpregnant women, and they have lower biological activity.
Prolactin is also synthesized in the brain including the hypothalamus, and in specialized
cells of the uterus, placenta, and breast, and in lymphocytes. In these areas it has
paracrine and autocrine functions.
There is a circadian pattern of prolactin secretion, which rises at night. The first peak
appears 60 to 90 minutes after the onset of slow-wave sleep, and subsequent peaks occur
later, after cycles of REM sleep. Stresses, including anesthesia, surgery, insulin-induced
hypoglycemia, fear, and mental tension, all cause prolactin increase. The purpose of
sleep- or stress-induced prolactin release is unknown.
Normal basal plasma concentrations of prolactin are about 10 ng/ml, and they are
similar in women and men. The half-life of the hormone is 20 minutes, and the daily
production is around 350 g. The kidney is a likely organ of prolactin degradation,
because patients with renal failure often have high plasma prolactin levels.

 Prolactin release is predominantly under tonic inhibition by dopamine derived from

hypothalamic dopaminergic neurons (cell bodies in the arcuate nucleus). These neurons
synthesize and secrete dopamine into the median eminence. This dopamine enters
capillaries that drain into the hypothalamic-hypophyseal portal vessels and deliver
dopamine directly and in high concentration to the
anterior pituitary, where it inhibits prolactin
secretion.
Interruption of the hypothalamic-pituitary tract
causes increased secretion of prolactin and sustained
lactation.
Dopaminergic inhibition of lactotroph release of
prolactin is mediated by dopaminergic Gi proteincoupled receptors, resulting in inhibition of adenylate
cyclase and inositol phosphate metabolism. In
addition, activation of the receptor modifies at least 5
different ion channels. In particular, dopamine
activates a potassium current that induces PM hyperpolarization while decreasing
voltage-activated calcium currents. Therefore, dopamine-induced inhibition of prolactin
secretion is a function of the inhibition of adenylate cyclase activity, activation of
voltage-sensitive potassium channels, and inhibition of voltage-sensitive calcium
channels.
Factors that alter prolactin secretion are summarized below.
Stimulatory Factors
Pregnancy (estrogen)
Breast-feeding
Sleep
Stress
TRH
Dopamine antagonists

Inhibitory Factors
Dopamine
Bromocriptine (dopamine
agonist)
Somatostatin
Prolactin (negative feedback)

Prolactin inhibits its own secretion by increasing the synthesis and secretion of dopamine
from the hypothalamus. This action constitutes negative feedback because stimulation of
dopamine secretion causes inhibition of prolactin secretion. Pregnancy and breastfeeding (suckling) are the most important stimuli for prolactin secretion.
In women. Prolactin release is affected by a large variety of stimuli provided by the
environment and the internal milieu, the most important being suckling, increased levels
of ovarian steroid hormones, primarily estrogen, and stress. The release of prolactin in
response to suckling is a classical neuroendocrine reflex. This surge in prolactin release
in response to a sucking stimulus is mediated by a decrease in the amount of dopamine
released by the hypothalamus, relieving the lactotroph from tonic inhibition. Estrogen
stimulates growth of the lactotrophs during pregnancy as well as prolactin gene
expression and release.

 Prolactin, in a supportive role with estrogen and progesterone, stimulates development

of the breasts, promotes milk secretion from the breasts during lactation, and suppresses
ovulation.
- Breast development. At puberty, prolactin, with estrogen and progesterone,
stimulates proliferation and branching of the mammary ducts. During pregnancy,
prolactin (again with estrogen and progesterone) stimulates growth and
development of the mammary alveoli, which will produce milk once parturition
occurs.
- Lactogenesis. The major action of
prolactin is stimulation of milk production
and secretion in response to suckling.
(Interestingly, pregnancy does not have to
occur for lactation to be possible; if there
is sufficient stimulation of the nipple,
prolactin is secreted and milk is
produced.) Prolactin stimulates milk
production by inducing the synthesis of
the components of milk, including lactose
(the carbohydrate of milk), casein (the
protein of milk), and lipids.
Although prolactin levels are very high during pregnancy, lactation does not
occur because the high levels of estrogen and progesterone down-regulate
prolactin receptors in the breast and block the action of prolactin. At parturition,
estrogen and progesterone levels drop precipitously, and their inhibitory actions
cease. Prolactin can then stimulate lactogenesis, and lactation can occur.
- Inhibition of ovulation. In females, prolactin inhibits ovulation by inhibiting
the synthesis and release of GnRH and therefore suppression of the female
reproductive cycle, which accounts for the reduced likelihood of another

conception during breastfeeding. [In males with high prolactin levels (e.g., due to
a prolactinoma), there is a parallel inhibitory effect on GnRH secretion and
spermatogenesis, resulting in infertility.]
Clinical correlation: The pathophysiology of prolactin can involve either a deficiency of prolactin, which
results in the inability to lactate, or an excess of prolactin, which causes galactorrhea (excessive milk
production) and decreased libido. Prolactin excess also causes failure to ovulate and amenorrhea because it
inhibits GnRH secretion.

Why do men produce prolactin? Prolactin plays an

important role in sex. In men, prolactin provides the body
with sexual gratification after sexual acts: The hormone
counteracts the effect of dopamine, which is
responsible for sexual arousal. This is thought to
cause the sexual refractory period.! At the
moment of orgasm, a sharp rise in prolactin levels
occur. As it is known to suppress other hormones
involved in reproduction, it is thought to suppress sexual
desire. That is why, after an orgasm, the person feels
satisfied and he doesn't need any further intercourse. The
amount of prolactin can be an indicator for the amount of
sexual satisfaction and relaxation. If men did not produce
prolactin, they might never be satisfied by intercourse and
always feel as if they need more. Unusually high amounts
are suspected to be responsible for impotence, loss of libido,!
mood!changes/depression,!weight!gain,!etc.

Dopamine!is!behind!a!lot!of!the!
desire!we!associate!with!eating!
and!sexual!intercourse.!!
Similarly,!all!addictive!drugs!
trigger!dopamine!(the!craving!
neurochemical)!to!stimulate!
the!pleasure/reward!center.!!So!
do!gambling,!shopping,!
overeating!and!other,!
seemingly!unrelated,!activities.!!
Go!shopping:!dopamine.!!
Smoke!a!cigarette:!dopamine.!!
Computer!games:!dopamine.!!
Heroin:!dopamine.!!Orgasm:!
dopamine.!!They!all!work!
somewhat!differently!on!the!
brain,!but!all!raise!your!
dopamine.!

OXYTOCIN
Oxytocin (OTC) is the other hormone released by the posterior pituitary. The primary
role of OTC is to eject milk from the lactating mammary gland and to facilitate uterine
contractions during labor.

 Both arginine vasopressin (or ADH) and OTC are synthesized in the cell bodies of
hypothalamic neurons. ADH originates largely in the supraoptic nucleus, and OTC
originates largely in the paraventricular nucleus. However, each hormone is also
synthesized in the alternate site.
OTC is a peptide of nine amino acids (a nonapeptide). The sequence is cysteine tyrosine - isoleucine - glutamine - asparagine - cysteine - proline - leucine - glycine. The
cysteine residues form a sulfur bridge. OTC has a molecular mass of 1007. OTC has a
half-life of typically about three minutes in the blood.

Since OTC has a role in

breastfeeding and in childbirth,
it is released in response to
breastfeeding and to cervical
and vaginal stimulation.
During breastfeeding, OTC
released by the neurohypophysis causes let-down
or expulsion of milk from the
mammary glands of the
breasts. Afferent neural
impulses are carried from
sensory receptors in the nipple
to the spinal cord, where they
ascend in the spinothalamic
tract. From relays in the
brainstem and midbrain, they
reach the paraventricular
nuclei of the hypothalamus.
From there, they trigger OTC
release from the
neurosecretory vesicles in the
posterior pituitary. As
suckling is continued, OTC
synthesis and its transfer down
the hypothalamic axon are also
stimulated.

 The unique effect of OTC is to cause contraction of the myoepithelial cells of the alveoli
of the mammary glands, in response to cries of the infant and the stimulus of suckling the
nipple. As a result, milk is forced from the alveoli into the ducts, from where it is
evacuated by the infant. The response is very rapid and milk flows within 1 minute.

OTC also has a powerful action on smooth muscle in the uterus. Formation of Ca++calmodulin complexes leads to activation of myosin light chain and to contraction of
smooth muscle cells. Rhythmic contractions of the myometrium are stimulated by very
small doses of OTC, which act by lowering slightly the threshold for membrane
depolarization. Larger doses lower the threshold still further, prevent repolarization and
spiking discharges, and induce a sustained tetanic contraction. Neither maternal plasma
OTC levels nor fetal OTC availability bears a consistent relationship to the progress of
labor during childbirth. Therefore, despite the ability to stimulate rhythmic uterine
contractions, secreted OTC seems to be more a contributing factor than an essential
hormone of human parturition. However, after delivery, it may play an important role in
the sustained contractions that help to maintain hemostasis after the placenta is
evacuated.
Clinical Correlation: Induction of Labor by Synthetic Oxytocin. Although a physiological role for
oxytocin in humans in induction or progression of labor is not well established, it stimulates strong uterine
contractions when used at high doses as a drug. Thus, synthetic oxytocin (sold as medication under the
trade names Pitocin and Syntocinon) is often used in clinical situations in which artificial induction or
stimulation of the progression of labor is medically required. As is generally the case for peptide drugs, it
is not effective orally due to cleavage in the gastrointestinal tract, and it is administered by intravenous
infusion.

In men. OTC and its receptor are found in the testis, epididymis, and prostate and thus
may assist in the movement of sperm, in ejaculation, and in the addition of seminal fluid
to the sperm. OTC plasma levels rise during male sexual activity, and they peak at
ejaculation. [OTC levels also rise during female sexual activity. The hormone and its
receptor are found in the ovary, and OTC assists in ovulation and in the termination of
the corpus luteum.] Recent studies have begun to investigate OTCs role in various
behaviors, including orgasm, social recognition, pair bonding, anxiety, trust, love, and
maternal behaviors.

STEROID SEX HORMONES

PATHWAY OF GONADAL STEROID SYNTHESIS
Both genders use the same pathway of steroid hormone
biosynthesis in gonadal tissue. The biosynthetic pathway of
steroid hormone in the gonads starts with cholesterol, and it
is essentially identical to that of the adrenal cortex. The
enzyme genes and characteristics, cofactor requirements
stimulators, and localizations are also the same as those for
the adrenal glands. More in situ cholesterol may be
synthesized from acetyl-CoA in the gonads than in the
adrenal glands.

P450-linked side chain

cleaving enzyme
(P450ssc) or 20,22desmolase, or
cholesterol desmolase!

The initial step in steroid hormone synthesis (steroidogenesis) is the conversion of

cholesterol to pregnenolone. Cholesterol used for steroid hormone synthesis can be
derived from the PM or from the cytosolic pool known as the steroidogenic cytoplasmic
pool. Cholesterol is released by the action of the enzyme cholesterol esterase.
Cholesterol is converted to pregnenolone by the action of the cytochrome P450 sidechain cleavage enzyme (P450SCC, also called 20,22-desmolase), which is present in the
inner mitochondrial membrane of all steroidogenic cells. For cholesterol to be a
substrate for P450SCC, free cholesterol must be transferred from the outer mitochondrial
membrane to the inner mitochondrial membrane, where the P450SCC enzyme is located.
This step is mediated by steroid acute regulatory protein.
Cholesterol is converted to pregnenolone by the removal of a 6-carbon fragment,
isocaproic acid the rate-limiting step in the gonads that is controlled by LH from the
anterior pituitary. This conversion of cholesterol to pregnenolone is the first step in a
sequence of enzymatic reactions involved in the synthesis of steroid hormones.

 Although two parallel synthetic pathways lead to testosterone, the delta 5 ( 5)

pathway from pregnenolone is favored. Oxidation of the A ring by the 3-oldehydrogenase-isomerase complex can take place at any point, from pregnenolone to
androstenediol. A small quantity of testosterone undergoes 5reduction to
dihydrotestosterone, and a further reduction of the 3-ketone position to 5
androstenediol also takes place within the testis.
Androgens are the obligate precursors of estrogens. The key step in conversion is
aromatization of the A ring; this reaction is heavily favored in the ovary and placenta.
The aromatase enzyme complex is a cytochrome P-450, localized in the endoplasmic
reticulum. It sequentially hydroxylates the 19-methyl group, oxidizes it to the aldehyde,
hydroxylates the 2 position, and then creates a 1-2 double bond by reduction. Following
these steps, the 19-carbon is removed by decarboxylation, and the characteristic benzene
ring is formed. Estradiol and estrone are formed from testosterone and androstenedione,
respectively. The two estrogens may also be interconverted by 17-hydroxysteroid
dehydrogenase.

PATHWAYS OF SYNTHESIS OF GONADAL STEROID HORMONES

20,22-desmolase (mitochondria)

3-ol-dehydrogenase
4,5
and -isomerase
17-hydroxylase

Estradiol!and!
progesterone!are!the!
major!secretory!
products!of!the!ovary.!

17-hydroxylase

3-ol-dehydrogenase
4,5
and -isomerase

17,20-desmolase

17,20-desmolase

3-ol-dehydrogenase
4,5
and -isomerase

aromatase

DHEA
17 -OH-steroid dehydrogenase

17 -OH-steroid dehydrogenase

3-ol-dehydrogenase
4,5
and -isomerase

17 -OH-steroid dehydrogenase

aromatase

5 -reductase
Testosterone!is!the!
major!secretory!
product!of!the!testis.!
3-reductase

DIT

TESTOSTERONE
Testosterone is synthesized and secreted by the Leydig cells of the testes.
The steroidogenic pathways in the testes are similar to those for the adrenal cortex, with
two important differences:
- The testes lack the enzymes 21-hydroxylase and 11-hydroxylase, and
therefore cannot synthesize glucocorticoids or mineralocorticoids.
- The testes have an additional enzyme, 17-hydroxysteroid dehydrogenase,
which converts androstenedione to testosterone. Thus, the androgenic endproduct of the testes is testosterone rather than DHEA and androstenedione (the
end-products of the adrenal cortex).
LH stimulates testosterone biosynthesis by
increasing mobilization and transport of cholesterol
into the steroidogenic pathway, an action that takes
place in minutes; as well as by stimulating gene
expression and activity of the steroidogenic enzymes,
a slower process that requires several hours.
Steroidogenic acute regulatory protein (also found in
the adrenal cortex) has a key role in the transfer of
cholesterol from the outer to the inner
mitochondrial membrane, the first step in steroid
hormone biosynthesis, for the conversion of
cholesterol to pregnenolone.
Pregnenolone is a key intermediate for all classes
of steroid hormones. After formation in the
mitochondria, it is transported to the SER, where the
remainder of the biosynthetic reactions takes place.
Pregnenolone can be converted to testosterone via
the delta 5 or delta 4 pathways. These refer to the
stage at which the double bond in ring B (position
5,6) is switched to ring A (position 3,4). The delta 5 intermediates include 17hydroxypregnenolone, DHEA, and androstenediol, while the delta 4 intermediates are
progesterone, 17-hydroxyprogesterone, and androstenedione. The delta 5 pathway is
the major route for testosterone production in the testis, whereas the delta 4 pathway
predominates in the ovary.
- Note that up until this last enzymatic reaction, the enzymatic steps involved in
testosterone synthesis are similar to those involved in androstenedione synthesis
by the adrenal glands. It is the activity of 17-hydroxysteroid dehydrogenase and
the enzymatic conversion of androstenediol to testosterone that are specific to
the testes.

 Although estrogens are only minor products of testicular steroidogenesis, they are
present as a normal circulating constituent in men. Androgens are converted to estrogens
by the action of the enzyme complex P450arom. The products of aromatization of
testosterone and androstenedione are estradiol and estrone, respectively. In men, there is
some aromatase activity in both Leydig and Sertoli cells, but aromatization mainly takes
place in peripheral tissues. Aromatization is a complex reaction involving removal of the
methyl group in position 19 and rearrangement of ring A into an unsaturated ring.
The action of LH on the Leydig cell is mediated through specific membrane receptors
of high affinity and low capacity. A single Leydig cell possesses about 15,000 LH
receptors. Occupancy of less than 5% of these is sufficient to elicit maximal
steroidogenic response.

The LH receptor is a single 93 kd glycoprotein that shares sequence homology with

other G protein-coupled receptors. Its structure reveals three functional domains: an
unusually large and glycosylated extracellular hormone-binding domain, a
transmembrane spanning domain that contains seven noncontiguous segments, and an
intracellular domain. Coupling of the receptor to a GS protein occurs on a loop of one of
the transmembrane segments.

 cAMP, produced in response to LH, activates protein kinase A. Although by itself quite
stable, cAMP is cleaved and inactivated by phosphodiesterase. LH stimulates
steroidogenesis by acting at two principal sites. One is phosphorylation of cholesterol
esterase, which releases cholesterol from its intracellular stores and transports it to the
mitochondria. The other is activation of the rate-limiting enzyme, mitochondrial
cytochrome P450scc, which yields pregnenolone from cholesterol.
Testosterone is not stored in the Leydig cells but diffuses into the blood immediately
after being synthesized. An adult man produces 6-7 mg/day of testosterone. This amount
slowly declines after age 50 to 4 mg/day by age 70-80. Hence, men do not undergo a
sudden cessation of sex steroid production upon aging, as women do during their
postmenopausal period.
Testosterone circulates bound to plasma proteins, with only 2-3% present as the free
hormone. About 30-40% is bound to albumin and the remainder to sex steroid-binding
globulin (SSBG), a 94 kd glycoprotein produced by the liver. SSBG binds both estradiol
and testosterone, with a higher binding affinity for testosterone. Because only free
(unbound) testosterone is biologically active, SSBG essentially functions as a reservoir
for the circulating hormone. The synthesis of SSBG is stimulated by estrogens ands
inhibited by androgens.

In some tissues, testosterone functions as a prohormone and is converted to the

biologically active derivatives dihydrotestosterone (DHT), estradiol, and
androstenedione. Skin, hair follicles, and most of the male reproductive tract contain an
active 5-reductase. The enzyme irreversibly catalyzes the reduction of the double
bond in ring A and generates DHT. Aromatization of testosterone to estradiol occurs in
fat, liver, skin, and brain tissue. Circulating levels of estrogens (estradiol plus estrone) in
men approach those of women during the early follicular phase, yet men are protected
from feminization as long as production of and tissue responsiveness to androgens are
normal. Androgens are metabolized in the liver to biological inactive derivatives suitable
for excretion by the kidney.

Clinical Correlation: 5-Reductase inhibitors such as finasteride (marketed as Proscar, Propecia, etc.)
block the conversion of testosterone to DHT and, therefore, block the production of active androgens in
some target tissues. Because the growth of the prostate gland and male pattern baldness depend on DHT
rather than testosterone, 5-reductase inhibitors can be used in low doses as a treatment for benign
prostatic hypertrophy and hair loss in males, and as treatment for prostatic cancer in high doses.

An androgen is a substance that stimulates growth of the male reproductive tract or

development of secondary sexual characteristics. Androgens have profound effects on
almost every body tissue: alterations in primary sex structures (e.g., testes and genital
tract); stimulation of secondary sex structures (i.e., accessory glands); development of
secondary sex characteristics responsible for masculine phenotypic expression; and
effects on psychology and sexual behavior. The relative potency ranking of androgens is
DHT > testosterone >> androstenedione > DHEA.
- Throughout adulthood, androgens are responsible for maintaining the structural
and functional integrity of all reproductive tissues.
Mediated by Testosterone
Differentiation of epididymis, vas deferens,
and seminal vesicles
Increased muscle mass
Pubertal growth spurt
Cessation of pubertal growth spurt
(epiphyseal closure)
Growth of penis and seminal vesicles
Deepening of voice
Spermatogenesis
Negative feedback on anterior pituitary
Libido

Mediated by Dihydrotestosterone
Differentiation of penis, scrotum, and
prostate
Male hair pattern
Male pattern baldness
Sebaceous gland activity
Growth of prostate

Androgens are responsible for:

- secondary sex characteristics and masculine phenotype
general body hair increases
beard, axillary and pubic hair
low-pitched voice, larynx enlarges
muscular build of the body (skeletal muscle)
skeletal growth, shoulders broaden
distribution of fat
- behavior
- anabolic effects
protein synthesis
in general, androgens promote protein synthesis and growth of
those tissues with androgen receptors
- epiphyseal plate
in the bones, testosterone (by way of aromatization to estradiol)
accelerates maturation of cartilage into bone, leading to closure of the
epiphyses and conclusion of growth

 Both functions of the testes, spermatogenesis and secretion of testosterone, are

controlled by the hypothalamic-pituitary axis. GnRH stimulates production of LH,
which, in turn, stimulates testosterone production from Leydig cells of the testes.
Testosterone acts via an intratesticular paracrine mechanism to reinforce the
spermatogenic effects of FSH in the Sertoli cells. Maturation of sperm requires both LH
and FSH. Inhibin (produced by the Sertoli cells) inhibits the secretion of FSH from the
anterior pituitary.

Clinical Correlation: Pubertal Growth Spurt.

A dramatic growth spurt occurs in both males and females during puberty. Somatic growth is primarily
controlled by the growth hormoneinsulin-like growth factor-1 axis. Increased LH and FSH secretion
at the beginning of puberty occurs together with increased hGH secretion. The increase of GH secretion is
caused by increasing estrogen levels in both sexes; in males, estrogen is produced from testosterone via the
enzyme aromatase. The highest rates of GH secretion occur during puberty because the strength of
negative feedback inhibition of GH secretion by IGF-1 is weak. After puberty, the rate of GH production
declines with age.
Sex steroids are responsible for closure of the epiphyseal growth plates in long bones. Early exposure to
sex steroids is likely to result in short adult stature. On average, men are 10-15 cm taller than women,
which can be attributed to two phenomena:
1. Growth during the pubertal growth spurt is more rapid in males than in females.
2. Puberty occurs later in males, allowing about 2 additional years of prepubertal growth before closure
of the epiphyseal plate is stimulated by sex steroids. Linear growth is completed in females by about age
17 and in males by about age 21.

Clinical Correlation: 5-reductase deficiency. 5-reductase deficiency is an autosomal recessive

disorder that results from the inability to convert testosterone to DHT. Differentiation of the internal
genitalia, which is testosterone-dependent, proceeds normally along the male line. The external genitalia,
which dependent on DHT, develop along the female line. Such individuals are often identified as females
at birth with bilateral inguinal masses, but undergo marked masculinization during puberty in response to
rising levels of testosterone.

Clinical Correlation: Testosterone DeficiencySigns of Andropause or Low T.

Testosterone deficiency Low T has the following signs and symptoms: decreased libido, sexual
dysfunction (impotence), infertility, small testes, and decreased muscle mass and strength. Associated
conditions include advanced age, obesity, liver disease, alcoholism, osteoporosis, rheumatoid arthritis,
AIDS, chronic renal failure, diabetes, and chronic anemia. In the United States, an estimated five million
men have the medical condition of testosterone deficiency (hypogonadism), which is approximately one in
twenty adult males.
The U.S. Government does not have a Recommended Dietary Allowance (RDA) for testosterone.
Forms of testosterone for human administration include injectable (such as testosterone cypionate or
testosterone enanthate in oil), oral Andriol, buccal Striant, transdermal skin patches, and transdermal
creams or gels such as AndroGel and Testim. Note: Important information about AndroGel. AndroGel
should not be used by a woman. Testosterone can cause birth defects in an unborn baby. A pregnant
woman should avoid coming into contact with AndroGel, or with a man's skin areas where AndroGel has
been applied.

Clinical Correlation: Anabolic steroids.

Anabolic steroids, or anabolic-androgenic steroids (AAS), are a class of steroid hormones related to the
hormone testosterone. They increase protein synthesis within cells, which results in the buildup of cellular
tissue (anabolism), especially in muscles. Anabolic steroids also have androgenic and virilizing properties,
including the development and maintenance of masculine characteristics such as the growth of the vocal
cords and body hair. The word anabolic comes from the Greek anabolein, "to build up", and the word
androgenic from the Greek andros, "man" + genein, "to produce".
Anabolic steroids were first isolated, identified and synthesized in the 1930s, and are now used
therapeutically in medicine to stimulate bone growth and appetite, induce male puberty, and treat chronic
wasting conditions, such as cancer and AIDS. The American College of Sports Medicine acknowledges
that AAS, in the presence of adequate diet, can contribute to increases in body weight, often as lean mass
increases, and that the gains in muscular strength achieved through high-intensity exercise and proper diet
can be additionally increased by the use of AAS in some individuals.
Some health risks can be produced by long-term use or excessive doses of anabolic steroids. These
effects include harmful changes in cholesterol levels (increased low-density lipoprotein and decreased
high-density lipoprotein), acne, high blood pressure, liver damage, and dangerous changes in the structure
of the left ventricle of the heart.
Ergogenic uses (i.e., any external influences that can positively affect physical or mental performance) for
anabolic steroids in sports and bodybuilding is controversial, because of their adverse effects and the
potential to gain an advantage conventionally considered "cheating." Their use is considered doping and
banned by all major sporting bodies. For many years the AAS have been by far the most detected doping
substances in IOC-accredited laboratories. In countries where AAS are controlled substances, there is often
a black market in which smuggled or even counterfeit drugs are sold to users.

SEXUAL DIFFERENTIATION
The process of sexual differentiation (i.e., the pattern of development of the gonads,
genital ducts, and external genitalia) produces the most fundamental and obvious
differences between the genders. However, during the first 5 weeks of gestation, the
gonads of males and females are indistinguishable and their genital tracts are unformed.
Between this stage of the indifferent gonad and that of the mature individual of either
gender, the process of sexual differentiation takes place.
SRY gene is the sex determining region of
the Y chromosome. SRY encodes the testisdetermining factor (TDF). SRY (TDF)
appears to initiate differentiation of the gonad
into a testis.

Anti-Mllerian hormone
causes the Mllerian ducts
to atrophy.

AHC, a gene on the X chromosome,

helps determine development of an
ovary.

The WnT gene stimulates

Mllerian duct development in
the absence of AMH.

REGULATION OF REPRODUCTION IN THE FEMALE

Gonadal hormones are tightly regulated by feedback mechanisms involving the
pituitary, hypothalamus, and gonads (and placenta).
An overview of the female reproductive system is shown in the diagram below. The
main reproductive hormones are shown in the boxes.

Ovarian sex steroids and neural monoamines (norepinephrine, epinephrine) may exert
stimulatory (+) or inhibitory (-) (or both) effects on the secretion of GnRH or pituitary
gonadotropins (or both). Note that dopamine inhibits prolactin release, but has no effect
on LH or FSH.

OVARIAN STEROIDS
The ovarian steroid hormones,
progesterone and 17-estradiol, are
synthesized by the ovarian follicles
through the combined functions of
the granulosa cells and the theca
cells. Virtually all steps in the
biosynthetic pathway are the same as
those for the adrenal cortex and the
testes. Recall that the adrenal cortex
produces all intermediates up to the
level of androstenedione, but
because it lacks the enzyme 17hydroxysteroid dehydrogenase, it
does not produce testosterone.
Recall also that the testes, having
17-hydroxysteroid dehydrogenase,
produce testosterone as their major
hormonal product. In the ovaries, all
steps in the biosynthetic pathway are
present, including aromatase, which
converts testosterone to 17-estradiol, the major ovarian estrogen.

These last steps of estrogen biosynthesis from

androgens include:
three successive hydroxylations of the 19-methyl
group of androgens,
followed by simultaneous elimination of the
methyl group as formate and aromatization of the
A-ring.

ESTROGEN STEROIDS
Estrogens are a group of steroid compounds, named for their importance in the estrus
cycle, functioning as the primary female sex hormones.
Estrogens are of three types: estrone (E1), estradiol (E2) and estriol (E3). At equal
concentrations, E2 has a stronger biological effect than E1, which is more powerful than
E3 (i.e., E2 > E1 > E3).

Natural estrogen compounds can contain any combination of these 3 types of estrogen;
the most common formulation is 10% estrone, 10% estradiol, and 80% estriol. Estriol is
thought to have a protective effect: estriol inhibits estrone and estradiol binding to the
estrogen receptor.
While estrogens are present in both men and women, they are usually present at
significantly higher levels in women of reproductive age. Estrogen functions include:
- promote the development of female secondary sex characteristics
enlargement of the breasts
body shape = narrow shoulders and broad hips
distribution of fat breasts and buttocks
no change in larynx
less body hair and more scalp hair
growth of pubic and axillary hair
-

(hair growth mainly due to androgensadrenals)

maturation of germ cells

development of the uterus
maintenance of the endometrium that will allow for blastocyst implantation
establishing the timing of ovulation
maintenance of pregnancy (along with the placenta)
mammary gland development and lactation

ESTROGEN STEROID PRODUCTION

Synthesis
Estrogen is produced primarily by developing follicles in the ovaries, the corpus luteum
and the placenta.
Some estrogens are also produced in smaller amounts by other tissues such as the liver,
adrenal glands and the breasts (these secondary sources are especially important in postmenopausal women).
Synthesis starts in the theca interna cells of the ovarian follicle, by the synthesis of
androstenedione from cholesterol; androstenedione then crosses the basement membrane
into the surrounding granulosa cells, where it is converted to estrone or estradiol, either
immediately or through testosterone. See the diagram below.
Dehydroepiandrosterone (DHEA) is an abundant sex steroid in women. DHEA is a
precursor steroid that can be converted to estrogens (estradiol) and androgens such as
testosterone and 5dihydrotestosterone.
Estrogens can be produced by the enzyme aromatase, which converts androgens such as
DHEA to estrogens, mainly estradiol and estrone.
Packaging
Estrogens, like the other steroids, diffuse out of cells they are made in. Estrogens are
not stored in vesicles. They are released immediately due to their lipid soluble nature.
Processing
The only processing that takes place are the conversion steps.
Release
Recall that steroids are released immediately due to their lipid solubility.
Degradation
Steroids are degraded by dehydrogenases and reductases in the liver and extrahepatic
sites. The inactivated sulfate esters and glucuronides are water-soluble and are excreted
in the urine.
E2 (Estradiol) Synthesis
In women of reproductive age, E2 is essentially produced by the enzymatic conversion
of androgens (androstenedione and testosterone). The androgens are produced by the
thecal cells under the influence of LH and their conversion to E2 occurs in the granulosa
cells of the follicle, through the enzyme aromatase. Aromatase activity depends on FSH
levels.
The steroidogenic output of the follicle requires cooperation between granulosa and
theca cells and coordination between FSH and LH. This is known as the two-hormone,
two-cell theory of ovarian steroidogenesis. The dashed line represents the inability of
granulosa cells to convert progesterone to androstenedione.

E1 (Estrone) Synthesis
In women of reproductive age, E1 is mainly produced from the enzymatic conversion of
androstenedione, which is secreted under the influence of LH by the thecal cells. The
aromatase activity depends on FSH. In menopausal women and in men, E1 and its
sulphate represent the main circulating estrogens. The biological function of E1 is
speculative, but it could be related to the regulatory effect that the conversion of E1 into
E2 has on the degree of estrogenization.

E3 (Estriol) Synthesis
In women of reproductive age, the very low concentrations of E3 are produced by
hepatic hydroxylation of E1 and E2. During pregnancy, E3 is produced in large
quantities from the fetal-placental unit. As 17-hydroxylase is lacking in the placenta and
3-hydroxydehydrogenase is absent in the fetus, the E3 production is dependent on a
fetal-placental collaboration. The E3 concentrations strongly increase during pregnancy,
indicating the fetal-placental cooperation. For this reason, the level of E3 has been used
to assess high-risk pregnancies. While the biological role of E3 is unknown, it is
considered the major estrogen of pregnancy. E3 has weak estrogenic properties in most
maternal organs, but increases uteroplacental blood flow.

 The placenta lacks enzymes needed to produce estrogens but has very high aromatase
activity to convert androgens to estrogens. The fetal adrenal glands supply the placenta
with weak androgens for conversion to estrogens. These glands produce
dehydroepiandrosterone sulphate (DHEA-S), most of which is converted to 16hydroxy-DHEA-S by the fetal liver. E3 is produced by the placenta from fetal 16-OHDHEA-S.

The fetus does not produce progesterone or estrogens, thereby avoiding exposure to
high concentrations of these steroids. Although the fetus produces large amounts of weak
adrenal androgens, masculinization of the female fetus does not occur because the
placenta acts as a large sink for fetal androgens.

Clinical Correlation: Selective Estrogen Receptor Modulators. The importance of understanding the
estrogen receptor (ER) has been greatly heightened by the discovery of selective estrogen receptor
modulators (SERMs), such as Raloxifene and Tamoxifen.
These pharmaceutically produced
nonsteroidal ER ligands have varied ER profiles. Tamoxifen antagonizes the action of estrogens on the
breast, but mimics the action of estrogens on the uterine endometrium. It is an excellent chemotherapeutic
agent for breast cancer, but it can also rarely produce endometrial cancer of the uterus. Raloxifene has
beneficial agonist effects on bone and serum lipids, but not on breast or endometrium. It is therefore a safe
treatment for osteoporosis, but its antagonist effects on the brain produce, as a side effect, the hot flashes
associated with estrogen deficiency.

Clinical Correlation: Menopause.

Menopause is a normal stage of lifethe physiologic effects of menopause can be viewed as a syndrome
of ovarian failure. Cessation of menstruation is a universal feature of menopause. Withdrawal of
estrogen is particularly important in accounting for the symptoms of menopause, which include insomnia,
hot flashes, variable degrees of vaginal atrophy, and decreased breast size. Estrogen deficiency causes
bone loss and increases the risk of osteoporosis.
During menopause, there is marked reduction of estrogen and progesterone concentrations. The loss of
ovarian function removes negative feedback on GnRH, producing high serum concentrations of FSH and
LH. FSH levels are particularly high in postmenopausal women because inhibins, which exert selective
negative feedback on FSH secretion, are no longer produced by the ovary.

Clinical Correlation: Hormone Replacement Therapy. Despite the proposed cardioprotective effects of
estrogen, the leading causes of death are similar in men and women: (1) cardiovascular disease, (2) cancer,
and (3) stroke. The Womans Health Initiative studied the effects of hormone replacement therapy
(combined use of estrogen and progesterone known as HRT) on postmenopausal women. The results
showed that, compared to women in the placebo group, women on HRT have higher rates of breast cancer,
coronary heart disease, stroke, and pulmonary embolism. The only benefits noted were reduced rates of hip
fracture and colon cancer. These findings from the Womans Health Initiative have contributed to the
controversy surrounding the use of combined estrogen and progestin.

PROGESTERONE STEROIDS
Two sex hormones play a role in the control of the menstrual cycle: estradiol and
progesterone. While estrogen peaks twice, during follicular growth and during the luteal
phase, progesterone remains virtually absent prior to ovulation, but becomes critical in
the luteal phase and during pregnancy. Many tests for ovulation check for the presence
of progesterone. These sex hormones come under the influence of the pituitary gland,
and both FSH and LH play necessary roles.
FSH stimulates immature follicles in the ovaries to grow. LH triggers ovulation. The
GnRH from the hypothalamus controls the pituitary, yet both the pituitary and
hypothalamus receive feedback from the follicle. After ovulation, the corpus luteum
which develops from the burst follicle and remains in the ovarysecretes both estradiol
and progesterone. Only if pregnancy occurs do hormones appear in order to suspend the
menstrual cycle, while production of estradiol and progesterone continues. Abnormal
hormonal regulation leads to disturbance in the menstrual cycle.
Functions of progesterone
Converts the endometrium to its secretory stage to prepare the uterus for implantation.
At the same time progesterone affects the vaginal epithelium and cervical mucus. If
pregnancy does not occur, progesterone levels will decrease, leading to menstruation.
Normal menstrual bleeding is progesterone withdrawal bleeding.
During implantation and gestation, progesterone appears to decrease the maternal
immune response to allow for the acceptance of the pregnancy.
Progesterone decreases contractility of the uterine smooth muscle by increasing the
RMP and preventing electrical coupling between myometrial cells. In addition,
progesterone decreases uptake of extracellular Ca++ required for contraction of
myometrial cells by down-regulating the expression of genes that encode subunits of
voltage-dependent calcium channels. Progesterone also prevents uterine contractions by
blocking the ability of estradiol to induce membrane expression of -adrenergic receptors
(-adrenergic activation causes contractions).
The fetus metabolizes placental progesterone in the production of adrenal
mineralocorticoids and glucosteroids.
A drop in progesterone levels is possibly one step that facilitates the onset of labor. In
addition, progesterone inhibits lactation during pregnancy. The fall in progesterone
levels following delivery is one of the triggers for milk production.
Progesterone is thermogenic, raising the core temperature. It reduces spasm and
relaxes smooth muscle. Bronchi are widened and mucus regulated. Progesterone
receptors are widely present in submucosal tissue. Progesterone acts as an antiinflammatory agent and regulates the immune response.

Progesterone synthesis
Progesterone, like all other steroid hormones, is
synthesized from pregnenolone, a derivative of cholesterol.
This conversion takes place in two steps. The 3-hydroxyl
group is converted to a keto group and the double bond is
moved to C-4 from C-5.

Progesterone is the precursor of aldosterone, and after conversion to 17hydroxyprogesterone to cortisol and androstenedione. Androstenedione can be converted
to testosterone, estrone, and estradiol. Progesterone is important for aldosterone
synthesis, as 17-hydroxyprogesterone is for cortisol, and androstenedione for sex
steroids.

HORMONAL PATTERNS DURING THE MENSTRUAL CYCLE

Plasma hormone levels throughout the menstrual cycle are seen in the graphs below.
Note the increases of estradiol from the dominant follicle and GnRH preceding ovulatory
surges of LH and FSH. The later broad peaks of progesterone and estradiol are produced
by corpus luteum secretion. The earlier inhibin B peak results from follicle production;
the later inhibin A peak results from corpus luteal production.

HORMONAL PATTERNS DURING PREGNANCY

The duration of pregnancy is, by convention, counted from the date of the last
menstrual period. Pregnancy lasts ~40 weeks from the onset of the last menstrual period,
or 38 weeks from the date of the last ovulation. Pregnancy is divided into three
trimesters, each of which corresponds to ~13 weeks. Hormone levels during pregnancy
are illustrated below.

The early peak of hCG rescues the corpus luteum, which is responsible for the early rise
in progesterone secretion. The placenta assumes production of progesterone by about
gestation week 9. Estriol secretion begins later because maturation of the fetal adrenal
glands, which supplies androgen precursors top the placenta, is necessary for estriol
production.

Placental Progesterone
The placenta is the main source of progesterone during pregnancy. From the luteal
phase to term, maternal progesterone levels rise six- to eight-fold. Although progesterone
originates almost entirely from the corpus luteum before 6 weeks gestation, its
production shifts to the placenta after the 7th week. Beyond 12 weeks, the placenta is
definitively the major source of progesterone.
While the placenta produces large amounts of progesterone, it has limited capacity to
synthesize cholesterol de novo. Maternal cholesterol in the form of low-density
lipoprotein (LDL) cholesterol is the principal source of precursor for the biosynthesis of
progesterone during pregnancy.
Progesterone functions to:
- promote endometrial stromal differentiation = decidualization;
- stimulates lobular-alveolar development of the mammary gland in preparation
for milk secretion but suppresses milk protein synthesis before parturition;
- inhibit smooth muscle contractility;
- decrease prostaglandin formation and increases the rate of prostaglandin
inactivation, which helps maintain myometrial quiescence and prevent the
onset of uterine contractions; and
- inhibit immune responses like those involved in graft rejection.
Like progesterone, during the first several weeks of gestation, and throughout the time
of the luteal-placental shift, 17-hydroxyprogesterone concentrations primarily reflect
the steroidogenic status of the corpus luteum. However, unlike progesterone, the ovaries
continue to be a significant source of 17-hydroxyprogesterone throughout pregnancy.
During the third trimester, the placenta also uses fetal D5-sulfoconjugated precursors to
secrete increasing amounts of 17-hydroxyprogesterone, and the placenta is the major
source of this hormone at term.

 The rise in 17-hydroxyprogesterone that begins at 32 weeks strongly correlates with

the fetal maturational processes known to begin at this time.
Clinical Correlation: Progesterone Replacement Therapy.
Progesterone is essential for bone formation, sex drive, and is a natural tranquilizer that helps women
cope with stress. Progesterone also helps provide protection to the uterus by keeping the endometrium
from thickening. With some women and some dosing schedules, the endometrial lining sheds from the
uterus through the vagina. Some women find this HRT-induced bleeding to be an unacceptable nuisance,
although with modern dosage regimens, the bleeding often dwindles or stops completely over time.
Combined Hormone Therapy. This replaces both estrogen and progesterone. Estrogen and
progesterone work in hundreds of areas in the body, including the brain, bones, breasts, blood vessels,
reproductive organs, urinary organs, skin, mucous membranes and the endocrine system. While estrogen
helps lighten many menopausal discomforts, it also causes growth of the lining of the uterus. This
thickening of the uterine lining is a risk factor for cancer. Taking a form of progesterone helps prevent
uterine cancer by protecting the uterine wall from the tissue build-up that can occur when estrogen is taken
alone. HRT also helps keep bones strong and the heart healthy. A combination of estrogen and
progesterone replacement is the most widely used therapy for women who have their uterus. There are two
forms of HRT: Natural hormones are medications that are chemically identical to what the body
produces. Synthetic hormones are medications that are similar but not identical to the hormones
naturally produced in the body.

Clinical Correlation: Selective Progesterone Receptor Modulators. A selective progesterone receptor

modulator (SPRM) is an agent that acts on the progesterone receptor. A characteristic that distinguishes
such substances from receptor full agonists (such as progesterone) and full antagonists (such as RU 486) is
that their action differs in different tissues (agonist in some while antagonist in others).
Mifepristone (RU-486) is a synthetic steroid used to prevent conception or induce medical abortions of
intrauterine pregnancies of up to 49 days gestation. [The name RU-486 is derived from an abbreviation for
the pharmaceutical company Roussel-Uclaf of France, which designed the drug, plus a serial number.] In
women, mifepristone at doses greater or equal to 1 mg/kg antagonizes the endometrial and myometrial
effects of progesterone. In medical abortion regimens, mifepristone blockade of progesterone receptors
directly causes endometrial decidual degeneration, cervical softening and dilatation, release of endogenous
prostaglandins and an increase in the sensitivity of the myometrium to the contractile effects of
prostaglandins. Mifepristone induced decidual breakdown indirectly leads to trophoblast detachment,
resulting in decreased syncytiotrophoblast production of hCG, which in turn causes decreased production
of progesterone by the corpus luteum (pregnancy is dependent on progesterone production by the corpus
luteum through the first 9 weeks of gestation--until placental progesterone production has increased enough
to take the place of corpus luteum progesterone production).

BIRTH CONTROL
Oral contraceptives contain combinations of estrogen and progesterone or
progesterone alone. The combination preparations exert contraceptive effects primarily
through negative feedback effects on the anterior pituitary. The contraceptive effect of
progesterone alone is based primarily on its effects on cervical mucus and tubal motility.
Oral birth control pills do not contain progesterone but a progestin.
[A progestin is a synthetic progestagen that has progestinic effects similar to progesterone. The
two most frequent uses of progestins are for hormonal contraception (either alone or with an
estrogen), and to prevent endometrial hyperplasia from unopposed estrogen in hormone
replacement therapy. Progestins are also used to treat secondary amenorrhea, dysfunctional
uterine bleeding and endometriosis, and as palliative treatment of endometrial cancer, renal cell
carcinoma, breast cancer, and prostate cancer.]

Hormonal contraception aims to disrupt the endocrinology of the menstrual cycle.

Clinical Correlation: Steroid Contraceptives. Combined oral contraceptive pills contain synthetic
estrogen (ethinyl estradiole or mestranol) together with one of several synthetic progestagens and produce a
range of effects on the female reproductive tract. Their main role of action is the inhibition of ovulation;
both estrogen and progestagen inhibit the ability of estrogen to produce a preovulatory surge of LH.
Estrogen also suppresses FSH and follicular development and ovulation. The degree of follicular activity
occurring during oral contraceptive use depends on the type and dose of steroid. The dose of estrogen used
in modern contraceptive preparations is likely to be the minimal amount (20 g ethinyl estradiol) that will
reliably suppress FSH enough to prevent the growth of an ovulatory follicle. Progestin-only pills produce
thick cervical mucus that inhibits sperm passage, and they also disrupt normal endometrial development
and inhibit gonadotropin production.
Method
Steroid contraceptives

Mechanism involved
Combination estrogen-progestin: constant
concentrations over a 21-day period followed by
7 days rest
Phasic estrogen-progestin: varying concentrations
throughout the 21-day period and 7 days of
placebo
Progestin only: constant progesterone dose daily

PARTURITION
Just as the maintenance of the pregnant state depends on a unique hormonal milieu, its
termination probably also depends on specific hormonal changes. However, the exact
mechanism by which parturition, or the process of giving birth, is initiated remains
unclear. Progesterone, estrogen, cortisol, relaxin, oxytocin, CRH, prostaglandins,
and catecholamines all influence the initiation and maintenance of labor and the final
uterine evacuation. The most dominant endocrine factor is a decrease in the
progesterone effect and an increase in the estrogen effect, commonly on the same targets.
Because species variations exist, it is difficult to extrapolate the results of animal studies directly to
humans, even if the studies have been performed in subhuman primates.

Myometrial quiescence
Myometrial activation
Myometrial gap junctions
Prostaglandin F2 and E production
Local oxytocin production
Oxytocin receptors
Cervical rigidity
Matrix metalloproteinase
Cervical ripening

Progesterone
+
NA
+
-

Estrogen
+
+
+
+
+
NA
+

+, Stimulates, -; inhibits; NA, not available.

The table above illustrates the effects of progesterone and estrogen on factors involved
in parturition. The diagram below illustrates current concepts of the endocrine regulation
of parturition.

Parturition, the
delivery of the fetus,
occurs ~40 weeks
after the onset of the
last menstrual period.

The fetus initiates signals that decrease the ratio of effective progesterone to estrogen in
the myometrium. This increases prostaglandin production, which abolishes uterine
quiescence and stimulates uterine contractions. OTC produced in the decidua and
placenta and a small maternal contributioncombined with markedly increased oxytocin
receptors as a result of the higher estrogen/progesterone ratiomay contribute to labor
but are not essential. However, OTC sustains uterine contractions after expulsion of the
fetus in order to minimize maternal loss of blood. Cortisol from the fetal adrenal,

stimulated by CRH and fetal ACTH, prepares the fetus to adapt to extrauterine life
successfully. Placental CRH is greatly augmented by mutual positive feedback effects
with cortisol and prostaglandins. CRH is also a myometrial stimulant. In addition, a
critical level of CRH may act as a placental alarm that triggers the process.
PARTURITION
The gestation period is approximately 40 weeks, and is measured from the first day of the
last menstrual period. Expulsion of the fetus, its placenta, and other membranes from the
uterus occurs during labor. This process is characterized by thinning and dilation of the
cervix and by strong regular contractions of the uterus. The four phases of uterine
activity during pregnancy and labor are:
Phase 0 is a long phase during which the uterus remains quiescent, under the influence of
progesterone. The peptide relaxin, secreted by the corpus luteum, assists in maintaining
the uterus in a relaxed state.
Phase&1!represents!activation!of!the!uterus!when!close!to!term.!!In!the!weeks!
preceding!parturition,!there!is!weak,!lowKfrequency!myometrial!activity!called!
contractures!(also!known!as!Braxton0Hicks!contractions!or!false!labor).!!In!phase!
1,!there!is!increased!myometrial!expression!of!receptors!for!oxytocin!and!excitatory!
prostaglandins&(PGE2,!PGF2 ).&&The!appearance!of!many!new!gap&junctions!
promotes!coordinated!contraction!of!the!myometrial!smooth!muscle!cells.!!These!
changes!are!associated!with!a!transition(from(contractures(to(true(contractions.!
!
Phase&2!is!the!dramatic!stage!of!labor!when!there!is!stimulation!of!the!uterus,!with!
increasing!oxytocin!and!prostaglandin!levels!inducing!true!uterine&contractions!
and!dilation&of&the&cervix.!!The(products(of(conception(are(delivered(at(parturition,(
ending(phase(2.!
!
In!phase&3,!the!uterus!returns!to!its!normal!size!with!the!assistance!of!sustained!
contractions!of!the!myometrium,!which!also!assists!in!blood!clotting.!

HYPOTHALAMUS-PITUITARY-ADRENAL/GONADAL AXIS
In addition to gonadal differentiation, normal sexual development also requires normal
development of the entire HPA/HPG axis, also referred as the reproductive axis. HPA
and HPG form a functional endocrine axis with hormonal regulation and feedback loops.
In contrast to the testes and adrenals, which produce hormones during fetal life, the
ovaries start to release steroid hormones when they are first stimulated by gonadotropins
at the onset of puberty.

PROTEINS VERSUS STEROIDS

Characteristic
Produced by
Size
Solubility
Receptors
Transported in blood

Proteins
Pituitary or
Syncytiotrophoblast
>5000 daltons
Hydrophobic
Membrane
Freely soluble

Steroids
Ovaries or Testes
~300 daltons
Hydrophilic
Intracellular
SHBG
(Sex Hormone Binding Protein)
or Albumin

REVIEW QUESTIONS
1. Which one of the following steroids is synthesized from cholesterol without being
hydroxylated by 17-hydroxylase?
A. Corticosterone
B. Cortisol
C. Testosterone
D. Estradiol
E. Androstenedione
2. Which step in testosterone synthesis is activated by LH?
A. Androstenedione to testosterone
B. Cholesterol to pregnenolone
C. Testosterone to DHT
3. Which steroidogenic enzyme is not present in the gonads?
A. 17-hydroxylase
B. 21-hydroxylase
C. cholesterol desmolase
4. Which hormone maintains the corpus luteum of pregnancy?
A. LH
B. hCG
C. Estradiol
D. Progesterone
5. Which of the following organs are needed to synthesize estrogen during the third
trimester of pregnancy?
A. Corpus luteum
B. Fetal adrenal cortex
C. Fetal liver
D. Maternal adrenal cortex
E. Maternal liver
F. Maternal ovaries
G. Placenta
6. In a genetic male with deficiency of 5-reductase, which of the following masculine
features are present?

A. Deepening of the voice

B. Epididymis
C. Male Hair distribution
D. Muscle mass
E. Testes
7. A man who has galactorrhea is found to have a prolactinoma. His physician treats
him with bromocriptine, which eliminates the galactorrhea. The basis for the therapeutic
action of bromocriptine is that it
A. antagonizes the action of prolactin on the breast.
B. enhances the action of prolactin on the breast.
C. inhibits prolactin release from the anterior pituitary.
D. inhibits prolactin release from the hypothalamus.
E. enhances the action of dopamine on the anterior pituitary.
8. Which of the following functions of the Sertoli cells mediates negative feedback
control of FSH secretion?
A. Synthesis of inhibin
B. Synthesis of testosterone
C. Aromatization of testosterone
D. Maintenance of the blood-testis barrier
9. Which of the following explains the suppression of lactation during pregnancy?
A. Blood prolactin levels are too low for milk production to occur
B. Human placental lactogen levels are too low for milk production to occur
C. The fetal adrenal gland does not produce sufficient estriol
D. Blood levels of estrogen and progesterone are high
E. The maternal anterior pituitary is suppressed
10. Which step in steroid hormone biosynthesis, if inhibited, blocks the production of all
androgenic compounds but does not block the production of glucocorticoids?
A. Cholesterol pregnenolone
B. Progesterone 11-deoxycorticosterone
C. 17-hydroxypregnenolene dehydroepiandrosterone
D. Testosterone estradiol
E. Testosterone dihydrotestosterone
11. Which step in steroid hormone biosynthesis occurs in the accessory sex target tissues
of the male and is catalyzed by 5-reductase?
A. Cholesterol pregnenolone
B. Progesterone 11-deoxycorticosterone
C. 17-hydroxypregnenolene dehydroepiandrosterone
D. Testosterone estradiol
E. Testosterone dihydrotestosterone
12. A 20-year-old male patient presents to the doctors office complaining about
continuous growth, lack of facial hair development, and smaller penis and testicles than
his college friends. Laboratory values include total testosterone 125 ng/dL and LH less
than 2 mU/mL. TSH and prolactin levels are normal. He has no history of medications,
drug use, or disease. GnRH stimulation test over a 7-day period produces elevations in
circulating levels of LH. Continued growth in this case is due to
A. increased estrogen production.

B. decreased inhibin release.

C. decreased testosterone production.
D. decreased sensitivity to LH stimulation.
13. Anabolic effects of androgens in bone are mediated by
A. testosterone-mediated increase in osteoblast apoptosis.
B. estradiol-mediated osteoblast proliferation.
C. DHT-mediated osteoclast activation.
D. testosterone-mediated decrease in osteoblast proliferation.
14. Which of the following can contribute to infertility in a young male body builder
taking anabolic steroids?
A. Decreasing circulating levels of DHT
B. Suppression of LH release from the pituitary
C. Decreasing testicular testosterone concentrations
D. Increasing muscle mass
15. Impaired activity of 5-reductase will affect
A. sexual differentiation and development.
B. male-pattern hair growth.
C. feedback regulation of GnRH release.
D. deepening of the voice.
16. During spermatogenesis, the spermatogonia undergo active mitosis
A. under the influence of FSH.
B. under the influence of testosterone.
C. under the influence of LH.
D. without specific hormonal stimulation.
17. The first enzymatic reaction, which is the rate-limiting step, in the production of
testosterone
A. occurs in the mitochondria.
B. occurs in the ribosomes.
C. involves aromatization.
D. generates progesterone as the immediate derivative.
18. Testosterone in the general circulation:
A. is bound to high-density lipoprotein.
B. is bound to androgen-binding protein.
C. is converted to DHT in the prostate.
D. is converted to 17-hydroxyprogesterone in the liver.
19. Stimulation of steroid synthesis by the mature Graafian follicle involves
A. binding of FSH to receptors on the theca cells.
B. binding of FSH receptors on the stroma.
C. activation of adenylate cyclase.
D. increased Ca++ influx via calcium-activated channels.
20. Granulosa cells cannot produce estradiol from cholesterol because they do not have
an active
A. 17-hydroxylase.
B. aromatase.
C. 5-reductase
D. sulfatase.

21. Treatment with a long-lasting progesterone preparation in an ovariectomized woman

will result in
A. decreased prolactin secretion.
B. increased FSH release.
C. induction of ovulation.
D. increase in basal body temperature by 0.5-1.0C.
22. The theca interna cells of the Graafian follicle are characterized by
A. their capacity for converting progesterone to androstenedione.
B. having both FSH and LH receptors.
C. exhibiting active aromatization of testosterone to estradiol.
D. their ability to produce inhibin.
23. A 30-year-old female patient arrives at your office because of missed menstrual
periods for 2 months. Her history indicates regular menstrual periods in the past. During
physical examination, you suspect that she may be pregnant. What laboratory values
would be compatible with your diagnosis?
A. Low plasma progesterone and high LH
B. High prolactin, low LH, and low progesterone
C. High urinary estradiol and low progesterone
D. High urinary hCG and high plasma progesterone
24. The effects of progesterone on the myometrium during pregnancy include
A. down-regulation of voltage-dependant calcium channel subunit expression.
B. increased estradiol-induced -adrenergic receptor expression.
C. decreased prostaglandin inactivation.
D. increased prostaglandin synthase activity.
25. A 34-year-old woman begins assisted fertility treatment to initiate ovulation. She is
treated with an agent that acts via GnRH receptors in the anterior pituitary. Which
treatment is most likely to be effective?
A. GnRH agonist given in pulsatile doses
B. GnRH agonist given continuously
C. GnRH receptor blocker given in pulsatile doses
D. GnRH receptor blocker given continuously
26. A 58-year-old man complained of loss of libido and reduced muscle strength. Serum
analysis showed reduced levels of testosterone. Reduced activity in which hormonetarget axis could account for his symptoms?
A. LH Leydig cell
B. FSH Leydig cell
C. LH Sertoli cell
D. FSH Sertoli cell
E. LH spermatogonium
F. FSH spermatogonium
27. A couple trying to conceive decided to use an ovulation predictor kit to indicate
when intercourse should occur to maximize the probability of pregnancy. If ovulation
occurs on day 15 of this womans menstrual cycle, when would implantation of a
successfully fertilized egg most likely occur in the uterus?
A. Days 16-17
B. Days 18-20
C. Days 21-22

D. Days 23-24
E. Days 25-26
28. A blood sample is taken from a 27-year-old woman for hormone analysis. The
sample contains low levels of FSH and LH and high levels of estrogen, progesterone, and
inhibin. Assuming a normal 28-day menstrual cycle, on which day of the womans
menstrual cycle was the blood sample taken?
A. Day 1
B. Day 7
C. Day 14
D. Day 21
E. Day 28
29. Which of the following enzymes or substrates does the fetus provide for the placenta
to produce estrogen?
A. Aromatase
B. DHEA-S
C. 21-Hydroxylase
D. Pregnenolone
E. Progesterone
F. 5-Reductase
G. Testosterone
30. A 23-year-old woman consulted her midwife 1 day after the uncomplicated delivery
of her first child. She was concerned about her lack of milk production and that she was
only producing a thin yellow fluid at the breast. The midwife reassured her that this
was normal and that milk production would begin within 1-2 days. Which hormone(s)
was suppressing her milk production?
A. Estrogen
B. Inhibin
C. Oxytocin
D. Progesterone
E. Prolactin
Answers: 1A, 2B, 3B, 4B, 5BCEG, 6ABDE, 7C, 8A, 9D, 10C, 11E, 12C, 13B, 14BC,
15AB, 16D, 17A, 18C, 19C, 20A, 21D, 22A, 23D, 24A, 25A, 26A, 27C, 28D, 29B,
30AD