Todays Goals

Describe mechanisms for development of male vs female

gonads
- Sex determination in mammals
Antiquity
Aristotle claimed that sex was determined by the heat
of the male partner during intercouse
The more heated the passion, the greater the
probability of male off-spring
Aristotle hypotesis : women were men whose
development was arrested to early
19th century : That factors favoring the storage of
energy and nutrients predisposed one to have female
offspring, whereas factors favoring the utilization of
energy and nutrients influenced one to have male
offspring
Chromosomal Sex Determination

Mammals: XX = female, XY = male

Avians: ZZ = male, ZW = female

Flies: 2X = female, 1X = male (Y = no role)

Bees, Ants: 2(n) = female, 1(n) = male

Chromosomal Sex Determination

Mammals
Mammalian Sex Determination and
Dev. Of the Gonad
The gonads are also derived from the intermediate
mesoderm
Form testes or ovaries
But before they choose - come from a common
precursor structure = Bipotential Gonad!
In the chromosomes - how is sex determination
achieved in mammals?
Mammalian Sex Determination Overview

Primary Sex Determination in Gonads

First form bipotential indifferent primordia gonad
Primary sex determination is chromosomal , not
influenced by environment
The egg forming ovaries or sperm forming testes
Most cases, XX ; Female, XY : Male
From intermediate mesoderm near mesonephros
Default is female, Y is crucial for SRY gene
Y chromosom carries a gene that encodes a testis
determining factor
XXXXXY : Male, XO : Female
Mammalian Sex Determination Overview
Secondary Sex Determination Elsewhere
Involves the sexual phenotype outside the gonads
A male : Penis, scrotum, seminal vesicles, and prostate
gland
A female : Vagina, clitoris, labia, cervix, uterus, oviduct
and mamary glands
Genitalia, mammaries, size, voice, musculature, hair
Endocrine, paracrine secretions from gonads
Absence of gonads produces female phenotype
The ovaries product estrogen mullerian duct
uterus, oviduct ,cervix, and upper end of the vagina
Testes form and secrete two major factors; Anti
mulerrian factor (AMF) destroy mullerian duct,
testosteron masculinizes the fetus.
Mammalian Gonads are Unique!
All other organ rudiments can only become 1 type of organ -
Lung rudiment - only forms a lung
But -the bipotential gonad that appears can become either ovary or
testes (genital ridge)
Appears around week 4 in human development, remains sexually
indifferent until week 7
The gonadal rudiments are paired regions of the intermediate
mesoderm : they form adjacent to the developing kidneys
The ventral portions of gonadal rudiments are composed of the
genital ridge epithelium.
These epithelial layers form the sex cord, the germ celss migrate
into the gonad during week 6, and are surrounded by the sex
cord. In both XY and XX gonads, the sex cords remain connected
to the surface epithelium
BASICS
Intermediate Mesoderm: The Urogenital System
Kidneys
Gonads
Respective Ducts
Basics!!
Development of Gonad
The gonad (testes and ovaries) are the organs that
produce cells (sperm and oocytes) . The gonad are
derived from three sources :
1. Mesothelium (mesodermal epithelium) lining the
posterior abdominal wall
2. Underlying mesenchyme (embryonic connective
tissue)
3. Primordial germ cell (earliest undifferianted sex
cells)
Basics!!!:
2 ducts form in sexually indifferent embryonic
stages
Has 2 developing ducts - Mllerian duct and
Wolffian duct
Based on hormonal interactions - one or the other
duct will degenerate, one will function, develop
further incorporating into reproductive tracts
Back to the larger view....
 Male gonad development
If the fetus XY, the sex cords continue to proliferate
through the eight week, extending deeply into the
connective tissue.
These cord fuse, forming a network of internal
(medullary) sex cords and at its most distal end, the
thinner rete testis, eventualy the sex cord-now called
testis cord-lose contact with the surface of epithelium and
become separated from it by a thick extracellular matrix,
the tunica albugenia.
The germ cell are found in the cords within testes. During
fetal life and childhood, the testis cord remain solid. At
puberty, however,the cords will holow out to form
seminiferous tubules and the germ cells will begin
differiante into sperm
Male gonad development
The cells of the seminerous tubule are called sertoli cells.
The sertoli cells of testis cord nurture the sperm and
secrete anti melerian duct hormone. The sperm are
transported form the inside of testis through the rete testis,
wich joints the efferent ducts
The eferent tubules are the remnants of the mesonephric
kidney, and they link the testis to the wolffian duct, which
used to be the collecting tube of the mesonepric kidney.
In Males, the wolffian ducts differentiates to become
epididymis and the vas deferens. The tube through which
the sperm pass into the uretra and out of the body, during
fetal development, the interstitial mesenchyme cells of the
testes differentiate into leydig cells which make testosteron
Female Gonad Developments
In the females, the germ celss will reside near the outer
surface of the gonad,. Unlike the sex cords in males,
which continue their proliferation, the initial sex cords
of xx gonads degenerate.
The epitelium soon produce a new set of sex cords,
which do not penetrate deeply into the mesenchyme,
but stay near the outer surface (cortex) of the organ,
thus they are called cortical sex cords.
These cords are split into clusters, which each clusters
surounding a germ cell. The germ cell will become the
ova, and the surounding cortical cell will diferiante
into granula cells.
Female Gonad Developments
The mesenchyme cells of the ovary differintiate into
thecal cells. Together the tecal and granulosa cells will
form the follicel that envelop the germ cells and and
secrete steroid hormones
Each follicle will contain a single germ cell. In females,
the mullerian duct remain intacts, and it differentiates
into the oviducts, uterus, cervix, and upper vagina.
The wolfian duct degenerates
Back to the larger view....
Back to the larger view...
Remember: Development of the vertebrate kidney

Bipotential gonad
starts as genital
ridge epithelium
in intermediate
mesoderm

Nephric duct
is the primitive
organizer:
Wolffian Duct

~ week 4 in humans
Differentiation of gonads in transverse section

Genital ridge forms as The primodial germ cell

originate in wall of the
epithelium mixed with
umbilical vesicle(from
interstitial mesenchyme epiblast and migrate
along the dorsal
mesentery of the
hindgut to the the
gonadal ridges

Mullerian duct
develops parallel
to Wolffian duct
Differentiation of the male gonad

Cell Fates:
- Epithelium becomes Sertoli cells
- Mesenchyme becomes Leydig cells
- Germ cells will become spermatogonia

- Sertoli cells line up to form solid cords

Ductal fates:
- Sertoli AMF degenerates Mullerian duct
- Leydig testosterone maintains Wolffian duct
- Testis cords will become seminiferous tubules
- Rete testis canals join testes cords to the
vas deferens and then epididymous.
Differentiation of the female gonad

Cell Fates:
- Epithelium becomes granulosa cells
- Mesenchyme becomes thecal cells
- Germ cells will become meiotic oogonia
- Together they form the follicles

Ductal fates:
- No AMF allows devo of Mullerian duct
- oviduct, uterus, cervix, upper vagina
- No testosterone degenerates Wolffian duct
Development of Male genital Ducts and Gland
Mesonephric ducts (Wolfian ducts) persist and are transformated into
the duct of epididymis.
Distal to epididymis mesonephric ducts acquires a thick investment of
smooth muscle and becomes to ductus deferens
Seminal Glands : Lateral outgrowths from caudal end of each
mesonephric duct. The part of mesonephric duct between the duct of
this gland and the urethra become ejaculation duct. (the secretion
nourish the sperms
Prostate : Multiple endodermal outgrowth arise from the prostatic
part of the urethra and grow into the surounding mesenchyme.
Secretions from the prostate contribute to the semen
Bulbourethral Glands : Pea sized structures developed from paired
outgrowths derived from the spongy part of urethra. Secretion
contribute to the semen
Development of Female Genital Ducts and Glands
The Paramesonephric ducts (Mullerian ducts) form
most of the female genital tract.
The uterine tubes developed from the unfused cranial
part of the paramesonephric ducts. The caudal, fused
portions of these ducts form the uterovaginal
primordium. Which gives rise to the uterus and the
superior portion of vagina
The Vaginal ephitelium is derived from the endoderm of
the urogenital sinus. The fibromuscullar wall of vagina
develops from surounding mesenchyme. Contact of the
ureterovaginal primordium with the urogenital sinus,
forming the sinus tubercle, induces the formation of
paired endodermal outhgrowths-sinovaginal bulbs. They
extend from the urogenital sinus to the caudal end of the
ureterovaginal primordium
Development of Female Genital Ducts and Glands
The sinovaginal bulbs fuse to form a vaginal plate. The
central cells of this plate break down, forming the lumen
of vagina.
Until late in fetal life, the lumen of the vagina is separated
from the cavity of the urogenital sinus by a membrane the
hymen formed by invagination of the posterior wall of the
urogenital sinus. The hymen rupturs during the perinatal
period
Female Auxiliary Genital Glands outgrowhts from the
urethra into surounding mesenchyme form the bilateral
mucus-secreting urethral glands and paraurethral
glands. Outgrowths from the urogenital sinus form the
greater vestibular glands in the lower one third of the
labia majora
Development of External Genitalia
Early in the fourth week, the proliferating mesenchyme
produces a genital tubercle primordium of penis or
clitoris.
In both sexes at the cranial end of the cloacal membrane.
Fgf 8 is involved in the signaling pathways in the early
development of the external genitalia
Labioscrotal swellings and urogenital folds soon
develop on each side of cloacal membrane. The genital
tubercle soon elongates to form primordial phallus
(penis or clitoris) . The urogenital membrane lies in
the floor of a median cleft, the urethral gr0ove, which
is bound by the urogenital folds.
In female fetuses, the urethra and vagina open into a
common cavity, the vestibule of vagina
Development of Male External Genitalia
Masculinization of the indifferent external genitalia is
induced by dihydrotestosteron from leydig cell
(testiscullar)
The primordial phallus enlarge and elongates to
become phenis, urogenital folds form the lateral wall of
the urethral groove on the ventral surface of the penis.
This groove is lined by a proliferation of endodermal cells
the urethral plate, which extends from the phallic portion
of urogenital sinus.
The urethral folds fuse with each other along the ventral
surface of penis to form spongy urethra.
The surface ectoderm fuses in the median plane of the
penis, forming the penile raphe and enclosing the spongy
uretra within the penis
At the tip of the glans penis, an ectodermal ingrowth,
extends toward the root of penis to meet spongy urethra
Development of Male External Genitalia
This cord canalizes and joint the previously formed
spongy urethra.
This juncture completes the terminal part of urethra
and moves the OUE to the tip of glans penis.
Corpora cavernosa and corpus spongiosum
developed from mesenchyme in the phallus.
The labioscrotal sweelings grow toward each other
and fuse to form scrotum.
Development of Female External Genitalia
Growth of the primordial phallus in the female fetus
gradually decreases as it becomes clitoris.
The clitoris develops in a similar way to the penis,
except that the urogenital folds do not fuse except
posteriorly, where they join to form the freenulum of
labia minora.
The unfused parts of the urogenital folds form the
labia minora .
The labioscrotal folds fuse posteriorly to form the
posterior labial commisure and anteriorly to form
the anterior labial commisure and mons pubis.
Most part of the labioscrotal folds remain unfused and
form two large folds of skin, the labia majora
Relocation of Testes and Ovaries
By 26 weeks, the testes have descend retroperitoneally from the
posterior abdominal wall to the deep inguinal rings. This change in
position occurs as the fetal pelvis enlarge and trunk of the embryo
elongates
Testiscular descent through ingunal canals and into the scrotum is
controled by testosteron. The gubernaculum guides testes through
inguinal canal and into the scrotum during the 26th weeks. When the
testes descend, they carry the ductus deferens decend. they are
ensheated by the fascial extensions of the abdominal wall :
The extension of the transversalis fascia becomes the internal
spermatic fascia
The extensions of the internal oblique muscle and fascia
become the cresmatic muscle and fascia
The extension of transversalis fascia becomes external
spermatic fascia
Within the scrotum, the testis projcts into the distal end of the
processus vaginalis tunica vaginalis (covers front & side testis)
Relocation of Testes and Ovaries
The ovaries also descend from the lumbar region of the
posterior abdominal wall and relocate to the pelvis;
however they do not pass from pelvis and enter the
inguinal canals.
The gubernaculum (fibrous cord) is attached to the
uterus near the attachment of the uterine tube.
The cranial part of the gubernaculum becomes the ovarian
ligament and the caudal parts forms the round ligament
of uterus.
The round ligament pass through inguinal canals and
terminate in the labia majora
The relatively small processus vaginalis in the female is
usually obliterated and it disappears long before birth, a
patent processus in a fetus is known as a vaginal process
of peritoneum (canal of Nuck)
Making decisions
Several human gene have been identified whose
function is necessary for normal sexual differentiation
The story starts in the genital ridge, which can become
either type of gonad. The genes encoding Wt 1, Sox9,
wnt 4,LHx9,Fgf9,GATA4 and Sf1 are expressed
Loss of function prevent the normal development
of gonad
Y chromosom (-) WNT4 , , Rspo Producing B
cathenin (+ feedback), Initiate the ovarian pathway
development, and prevent acumulation of Sox9 that
protein crucial for testis determination
Making decisions
If nucleus has a Y chromosome, the same set factor
activates the SRY gene. Sry protein binds to SOX9
promoter and elevates expression of this key gene in
the testis-determining pathway.
SOX 9 + FGF 9 block the ovary forming pathway,
blocking the function B-Catenin
BASICS!!! The male pathways : Make testes and dont
make ovaries.
The Female Pathways : Make ovaries and dont make
testes
The ovary pathway : Wnt4 and
R-spondin1
WNT 4 is expressed in genital ridges of both sexes,
later, undetectable in XY gonads . In transgenic xx
mouse lack the WNt4 gene, the ovary fails to form
properly.

RSPO1 critical in ovary formation, RSPO1 mutations

became males. RSPO1 +WNT4 B catenin. B catenin
critical in activating further ovarian development and
blocking syntesis Sox9
The testis Pathway (SRY)
The Y Chromosome sex determinant
In humans, the major gene for testis determination
resides on the short arm of the y cromosome.
Individuals born with the short but not the long arm
of Y chromosom are male, where as individuals born
with the long arm of the Y chromosm but not the short
arm are female.
Found in Normal XY Males, rare in XX Males, absents
form normal XX female and from many XY females
XX mice Transgenic for Sry Gene
The testis Pathway (SOX9)
Activate by SRY. SOX9 is an autosomal gene, In the genital
ridge SOX9 induces testis formation
XX humans who haves an extracopy of SOX9 developed as
males, even if they have no SRY gene
SRY (+) SOX9(-) mouse gonad cannot form testes
SOX9 protein has several function :
Blocks the ability of B catenin to induce ovary
formation.
It binds to cis regulatory regions of numerous gene
necessary for testis production.
SOX9 binds to the promoter site on the gene for the
AMF.
SOX9 promotes the expresions of the gene
encoding FGF9, establishing testis development.
The testis Pathway (FGF9)
When the gene for FGF is knocked out in mice, the
homozygous mutants are almost all female.
FGF9, whose expresions is activated by Sox9 plays
several roles
Causes proliferation of the sertoli cells precusors
and stimulates their differentiation
Activates the migrations of endothelial cells from
the adjacent mesonephros into the xy gonad, these
endotelial cells form the artery of testis and form
testis cords.
The testis Pathway (SF1)
A critical link between SRY and the male development
pathway
The transcription factor steroidigenic factor1 (SF1) is
necessary to make bipotensial gonad.
SRY maintains SF1 expresions. SF1 protein appears to
be active in masculinizing both the leydig & sertoli
cells. SF1+SRY > SOX9
Elevates levels of AMF
In the leydig cells, activate genes encoding enzymes
that make testosterone.
How it works
In an XX or any other combo lacking a Y chromosome
(XO, XXX)
No Y present, gonad develops into ovaries
Ovaries release estrogen
This enables development of Mllerian duct
Forms uterus, oviducts, upper end of the vagina

Female is the default sex

How it works
If Y chromosome is present
SRY gene is expressed
Testes determining factor is produced
Induces formation of testes
Secrete 2 hormones:
Anti-Mllerian Hormone

Testosterone

Causes Mllerian Duct to degenerate

Allows Wolffian Duct to connect to developing male gonad

Secondary Sex Determination

Gonadal determination is done by ~20 wks

True hermaphroditism is very rare in humans
Can result from translocation of Y onto X

Have two major temphoral phases

First phase: organogenesis second : puberty
The dominant influence is gonadal steroid hormones
In females the mullerian ducts persists, through the estrogen
actions differientiate to become uterus,cervix, oviduct and
upper vagina. The genital tubercle clitoris, The labioscrotal
folds labia
In male, testosteron causes the wolffian ducts to differentiate
into the epididymis, vas deferens, and seminal vesicles,
genital tubercles penis, labioscrotal scrotum
Unusual Cases in Sex
Determination
Androgen Insensitivity -
person is XY, produces SRY gene product, and thus
testosterone
No receptor for testosterone!
Have testes, but also make and respond to estrogen
Thus phenotype on the outside is female (secondary sex
characteristics) and they have a uterus and oviducts
Androgen insensitivity syndrome
These xy individuals have
the sry genes and thus
have testes that make
testosterone and AMF.
They have a mutation in
the gene encoding the
androgen receptor
protein that binds
testosterone. Cannot
respond to the
testosterone mad by their
testes

They can respond to estrogen

Congenital adrenal hyperplasia in females
Female pseudohermaproditism (in which the gonadal
sex is female but the person is outwardly male) can be
caused by the overproduction of androgen in the ovary

Androgens accumulate because of a mutation in the

adrenal enzyme to make corticosteroids (testosteron
like steroid)

Allows male secondary characteristics in a gonadal

female
Testosterone must be converted to dihydrotestosterone to finish male system.

Failure to convert causes

external genitalia to fail to
form prior to puberty

The children are often

raised as girls until the
high testosterone levels
of puberty finish the
job!
 ANTI-MULLERIAN FACTOR (AMF)
Member of TGF B Family of growth and differentiation
factors, secreted from the fetal sertoli cells and causes th
degeneration of the mullerian duct, Induces apoptosis in
ducts epithelium and breaks down the basal lamina duct
ESTROGEN
Needed for complete postnatal development of both
mullerian and wolfian duct.
Necesarry for fertility in both male and female
Induces the diff of mulerian duct become uterus
oviducts, cervix and upper vagina
Unusual Cases in Sex
Determination
XO individuals
Initially develop ovaries, but they degenerate
Appearance is female, have female genital tract, due to
prenatal estrogen exposure but sterile
Unusual Cases in Sex
Determination
Pseudohermaphroditism
One type of gonad, but opposite secondary sex
characteristics (as in androgen insensitivity)
Female pseudohermaphroditism - gonads are female,
but outwardly male - can occur (overproduction of
androgen hormones)
True Hermaphrodites
Contain both male and female gonadal tissue
Results from abnormalities in primary sex
determination (X/Y system)
Translocations of Y chromosome INTO X
chromosomes
Gynandromorph finch with ZZ (male) cells on its right side and
ZW (female) cells on its left side

True
Hermaphrodite

Weve also
seen
them in
worms...
Sex Determination in the Brain

Interestingly, the brain doesnt wait for the gonads

like most tissues do
More than 50 genes are expressed in a sexually
dimorphic pattern prior to gonad differentiation
These include SRY (active in the substansia nigra of
male hypothalamus)
Song centers in male birds can form w/o testosterone
Gonadal hormones refine an already present sex
Temperature-dependent sex determination in reptiles: American alligator,
red-eared slider turtle, and alligator snapping turtle
Increased estrogen can override temperature

Percentage of males crashes

We put key things into environment

Atrazine weed killer

Increased estrogen can override temperature
Other Environmental Sex Determinants

Physical position
Bonellia
Larva drops to sea floor - female
Larva finds proboscis of adult female male

Population conditions
Goby fish
School is one male, many females
If male is lost, top females race to change
Most aggressive becomes dominant male
Figure 14.25 Sex determination in
Bonellia viridis
Note:
The mammalian X/Y system of sex determination
is not used in all species
For most species, other systems are used
In Drosophila a ratio system is used
Ratio between number of X chromosomes and number
of autosomes
if there is 1X, fly is male, if 2X fly is female
NOT dependent on Y
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