SEX DETERMINATION

IN MAMMALS

LIPO MARIA CARMELA R.

 Chromosomal Sex Determination in Mammals

1. Primary Sex Determination

2. Secondary Sex Determination

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 Chromosomal Sex Determination in Mammals:
Primary Sex Determination

• Primary sex determination is the determination of the gonads.

• In mammals, primary sex determination is strictly chromosomal.
• The female is XX and the male is XY.
• Since the female is XX, each of her eggs has a single X chromosome.
• The male, being XY, can generate two types of sperm:
1. half bear the X chromosome,
2. half the Y.
• Active and Gene-directed processes.
• Both diverge from a common precursor, the bipotential gonad.

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 Chromosomal Sex Determination in Mammals:
secondary Sex Determination

• Secondary sex determination affects the bodily phenotype outside the gonads.
• Each sex has a sex specific size, vocal cartilage, and musculature.
• These secondary sex characteristics are usually determined by hormones secreted from
the gonads.
• X Chromosome: ovaries estrogen Müllerian duct female phenotype
• Y chromosome: testes two major hormones:
1. anti-Müllerian duct hormone (AMH) destroy Müllerian duct
2. testosterone male phenotype
• The body has a female phenotype unless changed by the two hormones secreted.

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Figure 1. General Scheme of Mammalian Sex Determination

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 The developing gonads

• The gonadal rudiment has two normal options when differentiates, it can develop into
either an ovary or a testis.
• Bipotential (indifferent) stage, during which time it has neither female nor male
characteristics.

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The developing gonads
testis

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The developing gonads
testis

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 The developing gonads
testis
SERTOLI CELLS
➢ the cells of the seminiferous tubule
➢ Nurture the sperm and secrete anti-Müllerian duct hormone.
RETE TESTIS
➢ The sperm are transported from the inside of the testis through the rete
testis
➢ Joins the efferent ducts.
EFFERENT DUCTS
• the remnants of the mesonephric kidney, and
• they link the testis to the Wolffian duct, which used to be the
collecting tube of the mesonephric kidney.
WOLFFIAN DUCT
➢ Become the epididymis and the vas deferens,
INTERSTITIAL MESENCHYME CELLS
• Differentiate into Leydig cells, which make testosterone. 9
 The developing gonads
ovary

• Germ cells ova

• cortical sex cords granulosa cells.
• The mesenchyme cells thecal cells.
• The thecal and granulosa cells follicles

• In females, the Müllerian duct remains intact,

and it differentiates into the oviducts, uterus,
cervix, and upper vagina. The Wolffian duct,
deprived of testosterone, degenerates.

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 Mechanisms of mammalian sex determination:
Sry: the Y chromosome sex determinant

• Found on the sex determining region of the Y chromosome.

• Produces testis determining factor.

MODES OF ACTION:
1. Bipotential gonads Testes.
2. Directly: Epithelium Sertoli cell.
3. Indirectly: Mesonephric cell Epithelium Sertoli Cell.

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 Mechanisms of mammalian sex determination:
Sox9: autosomal sex reversal

• Encodes a transcription factor that also contains a HMG box.

• XX humans who have an extra copy of SOX9 develop as males (no SRY gene).
• Having only one functional copy of this gene develops as female or hermaphrodites.
• Campomelic dysplasia, a disease involving numerous skeletal and organ systems.
• Expression is seen in the same genital ridge cells as Sry, and is expressed just slightly
after Sry expression.
• Activated by its relative, Sry.

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 Mechanisms of mammalian sex determination:
Sf1: the link between sry and the male development pathways

• Sf1 (Steroidogenic factor 1) is a transcription factor that may be directly or indirectly

activated by SRY.
• Sf1 is necessary to make the bipotential gonad; the Sf1 gene stays on in the developing
testis.
• In the Leydig cells, Sf1 activates the genes encoding the enzymes that make testosterone.
• In the Sertoli cells, Sf1, together with Sox9, is needed to elevate the levels of AMH
transcription.
• SRY gene activates Sf1 and then Sf1 protein activates the components of the male
differentiation pathway.

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 Mechanisms of mammalian sex determination:
Dax1: a potential ovary-determining gene on the x chromosome

• Having 2 copies of Dax1 on the active X chromosome

of the XY individual, the SRY signal would be
reversed.
• Expressed in the same genital ridge as the SRY gene.
• Dax1 antagonize the function of SRY, and down-
regulates Sf1 expression.
• Dax1 is a gene that is involved in ovary determination.

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 Mechanisms of mammalian sex determination:
Wnt4: a potential ovary-determining gene on an autosome

• The Wnt4 gene is expressed in the mouse genital ridge

while it is still in its bipotential stage.
• Wnt4 is undetectable in XY gonads (which become testes),
whereas it is maintained in XX gonads as they begin to
form ovaries.
• In transgenic XX mice that lack the Wnt4 genes, the ovary
fails to form properly, and its cells express testis-specific
markers, including AMH- and testosterone-producing
enzymes.
• Sry may form testes by repressing Wnt4 expression in the
genital ridge, as well as by promoting Sf1.

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 Secondary sex determination:
hormonal regulation of the sexual phenotype

• The development of the female and male phenotypes in response to hormones secreted by
the ovaries and testes.
• Both female and male secondary sex determination have two major temporal phases.
1. The first occurs within the embryo during organogenesis;
2. The second occurs during adolescence.
Two testicular hormones:
1. AMH- made by the Sertoli cells that causes the degeneration of the Müllerian duct.
2. STEROID TESTOSTERONE- secreted from Leydig cells. Causes the Wolffian duct to
differentiate into male phenotype, and it causes the urogenital swellings to develop into
the scrotum and penis.
• The existence of these two independent systems of masculinization is demonstrated by
people having androgen insensitivity syndrome.

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 Testosterone and dihydrotestosterone

• Testosterone is responsible for promoting the formation of the male reproductive

structures (the epididymis, seminal vesicles, and vas deferens) that develop from the
Wolffian duct primordium.
• 5a-dihydrotestosterone masculinize the male urethra, prostate, penis, or scrotum.
• Testosterone is converted to 5a- dihydrotestosterone with the help of the 5a-keterosteroid
reductase 2 in the urogenital sinus and swellings, but not in the Wolffian duct.
• The formation of the external genitalia is under the control of dihydrotestosterone,
whereas Wolffian duct differentiation is controlled by testosterone itself.

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 Anti-Mullerian duct hormone

• Anti-Müllerian duct hormone (AMH), is a 560-amino acid glycoprotein secreted from

the Sertoli cells.
• Cause the degeneration of the Müllerian duct.
• AMH is thought to bind to the mesenchyme cells surrounding the Müllerian duct and to
cause these cells to secrete a paracrine factor that induces apoptosis in the Müllerian duct
epithelium.

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 estrogen

• Estrogen is needed for the complete development of both the Müllerian and the Wolffian
ducts.
• In females: Induce the differentiation of the Müllerian duct into its various components:
the uterus, oviducts, and cervix.
• The extreme sensitivity of the Müllerian duct to estrogenic compounds is demonstrated by
the teratogenic effects of diethylstilbesterol (DES), a powerful synthetic estrogen that
can cause infertility by changing the patterning of the Müllerian duct.
• In males: estrogen is actually needed for fertility.
• While blood concentrations of estrogen are higher in females than in males, the
concentration of estrogen in the rete testis is even higher than that in female blood.

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 estrogen

• Androgen insensitivity syndrome is one of several conditions called

pseudohermaphroditism.
• In a pseudohermaphrodite, there is only one type of gonad, but the secondary sex
characteristics differ from what would be expected from the gonadal sex.
• Male pseudohermaphroditism can be caused by mutations in the androgen receptor or by
mutations affecting testosterone synthesis.
• Female pseudohermaphroditism can be caused by an overproduction of testosterone.
• True hermaphrodites contain both male and female gonadal tissue.
• Mammalian true hermaphrodites result from abnormalities of primary sex determination.
This abnormalities occur when the Y chromosome is translocated to the X chromosome.
• Translocated X chromosome is inactivated: SRY gene is turned off.
• Translocated X chromosome is not inactivated: SRY gene is on.

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 Sex determination and behaviors:
Organization/Activation Hypothesis

• Organization/activation hypothesis is the idea that sex hormones may act during the
fetal or neonatal stage of a mammal's life to organize the nervous system in a sex-specific
manner.
• Estradiol a hormone responsible for determining the male brain pattern is.
• P450 aromatase the enzyme that converts testosterone into estradiol. This conversion
occurs in the hypothalamus and limbic system two areas of the brain known to regulate
hormone secretion and reproductive behavior.
• a-fetoprotein bind and inactivate estrogen, but not testosterone. Is made in the fetal liver
and becomes a major component of the fetal blood and cerebrospinal fluid.

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 Sex determination and behaviors:
Male homosexuality

• Simon LeVay proposed that part of the anterior

hypothalamus of homosexual men has the anatomical form
typical of women rather than of heterosexual men.
• The interstitial nuclei (neuron clusters) of the anterior
hypothalamus (INAH) were divided into four regions.
Three of them showed no signs of sexual dimorphism.
However, one of them, INAH3, showed a statistically
• LeVay claimed, "suggests that sexual orientation has a
biological substrate."

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 Sex determination and behaviors:
Male homosexuality

CRITICISMS:
• First, the data are from populations, not individuals.
• Second, the "heterosexual men" were not necessarily heterosexual, nor were the
"homosexual men" necessarily homosexual; the brains came from corpses of people
whose sexual preferences were not known.
• Third, the brains of the "homosexual men" were taken from patients who had died of
AIDS.
• Fourth, because the study was done on the brains of dead subjects, one cannot infer cause
and effect.
• Fifth, there is no evidence that the difference has anything to do with sexuality.
• Sixth, these studies do not indicate when such differences emerge.

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SEX DETERMINATION
IN DROSOPHILIA

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 THE SEXUAL DEVELOPMENT PATHWAY

• In Drosophila, sex determination is achieved by a balance of female determinants on the

X chromosome and male determinants on the autosomes.
• MALE: One X chromosome in a diploid cell (1X:2A)
• FEMALE: Two X chromosomes in a diploid cell (2X:2A)
• XO Drosophila are sterile males.
• Y chromosome is not involved in determining sex. It contains genes active in forming
sperm in adults.

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 THE SEXUAL DEVELOPMENT PATHWAY

• Gynandromorphs animals are insects in which certain regions of the body are male and
other regions are female.
• Happens when an X chromosome is lost from one embryonic nucleus. The cells
descended from that cell, instead of being XX (female), are XO (male).
• The XO cells display male characteristics, whereas the XX cells display female traits.

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 THE SEXUAL DEVELOPMENT PATHWAY

• Loss-of-function mutations in most of these genes Sex-lethal (Sxl), transformer (tra),

and transformer-2 (tra2) transform XX individuals into males.
• Homozygosity of the intersex (ix) gene causes XX flies to develop an intersex phenotype
having portions of male and female tissue in the same organ.
• The doublesex (dsx) gene is important for the sexual differentiation of both sexes. If dsx
is absent, both XX and XY flies turn into intersexes.

The positioning of these genes is based on

(1) the interpretation of genetic crosses resulting in flies bearing two or more of these
mutations and
(2) the determination of what happens when there is a complete absence of the products of
one of these genes.

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 The sex lethal gene

First Phase: Interpreting the x:a Ratio

• High values of the X:A ratio are responsible for activating the feminizing switch gene
Sex-lethal (Sxl).
• In XY cells, Sxl remains inactive during the early stages of development.
• Sxl is activated during the first 2 hours after fertilization, and this gene transcribes a
particular embryonic type of Sxl mRNA that is found for only about 2 hours more.
• Once activated, the Sxl gene remains active because its protein product is able to bind to
and activate its own promoter.

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 The sex lethal gene

NUMERATOR PROTEINS
• Stimulates the female-specific activation of Sxl.
• These numerator proteins include Sisterless-a and Sisterless-b.
• These proteins bind to the "early" promoter of the Sxl gene to promote its transcription
shortly after fertilization.

DENOMINATOR PROTEINS
• Are autosomally encoded proteins such as Deadpan and Extramacrochaetae.
• These proteins block the binding or activity of the numerator proteins.
• The denominator proteins may actually be able to form inactive heterodimers with the
numerator proteins.

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 The sex lethal gene

There are 3 genes involved in the sex determination of Drosophila

1. Sxl gene (Sex lethal gene) – Splicing repressor

2. Tra gene (Transformer gene) - Activates splicing
3. Dsx gene (Doublesex gene) – Regulates sex expression

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 The sex lethal gene

MALE DROSOPHILA

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The transformer genes

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 Environmental sex determination:
TEMPERATURE-DEPENDENT

TURTLES.
• Eggs incubated at low temperatures (22-27°C) produce male.
• whereas eggs incubated at higher temperatures (30°C and above) produce female.
• Sex determination in reptiles (and birds) is hormone-dependent.
BIRDS AND REPTILES
• Estrogen can override temperature and induce ovarian differentiation even at
masculinizing temperature.
• Injecting eggs with inhibitors of estrogen synthesis will produce male offspring, even if
the eggs are incubated at temperature that usually produce females.

Aromatase, which can convert testosterone into estrogen is important in temperature

dependent sex determination.

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 Environmental sex determination:
LOCATION-DEPENDENT SEX DETERMINATION IN BONELLIA AND CREPIDULA

Bonella Crepidula fornicate

• Ocean floor : 10-cm-long female.
• Attracted to a female's proboscis it
tube female’s body: 3-mm-long
male. It becomes a sperm-producing
symbiont of the female.
• Young individuals are always male.
• Next phase is the degeneration of the male
reproductive system.
• The next phase can be either male or female,
depending on the animal's position in the mound.
• If the snail is attached to a female, it will become
male.
• If such a snail is removed from its attachment, it
will become female
• The presence of large numbers of males will cause
some of the males to become females.

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THANKS FOR LISTENING,SIS!

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