Developmental Biology

XL 138
Week 10
Development of the Endoderm
Anterior-Posterior
specification of the
gastrointestinal tract
Reciprocal Induction
Simultaneous
Anterior-Posterior
specification of
both endoderm
and mesoderm
https://www.youtube.com/watch?v=cBSyOgjTGVU
https://www.youtube.com/watch?v=3JAqrRnKFHo
 Levels of Sexual Development
• There are 3 levels to sexual development:
– chromosomal sex: presence or absence of the Y chromosome
– gonadal sex (primary sex determination): whether the gonads
develop as testes or ovaries depends on the presence or absence
of the SRY gene, usually found on the Y chromsome
– phenotypic sex (secondary sex determination): all of the internal
and external structures develop along male or female lines
depending on which hormones are secreted by the gonads.
• Phenotypic sex also has a couple of distinct systems: the
internal ducts, and the external genitalia
• Two important times: pre-natal development and puberty
 Chromosomal Sex Determination
• We have 46 chromosomes: 23 pairs,
one set from each parent.
• One pair of chromosomes is the sex
chromosomes, X and Y.
– the other chromosomes just have
numbers: 1-22.
• A person with 2 X chromosomes
(46,XX) is female, and a person with
an X and a Y (46,XY) is male.

Human karyotype: chromsomes

stained to show bands, from a male
 The SRY Gene
• How the Y chromosome determines sex.
• The SRY gene, located on the Y chromosome, is the
primary determinant of sexual development.
– That is, if a developing embryo has a functional SRY
gene in its cells, it will develop as a male. And, if
there is no functional SRY, the embryo develops as
female.
• Although the SRY gene is usually on the Y
chromosome, it occasionally gets transferred to the
X.
– this leads to 46,XX males
• Also, sometimes the SRY gene is inactivated by
mutation.
– Leading to 46,XY females (Swyer syndrome)
– it is also possible to have a partially inactive SRY gene,
leading to ambiguous genitalia
 Early Gonad Development

• Before 6-7 weeks of

development, the gonad
is indifferent: neither
male nor female.
• It develops from the
same tissue as the
kidneys and adrenal
glands.
• Also developing by this
time: 2 sets of ducts that
will eventually lead to the
outside world.
– Wolffian ducts = male
– Mullerian ducts =
female
 Gonad Differentiation
• If SRY is present in the
indifferent gonad at 6
weeks, it gets activated.
This in turn activates other
genes, and the indifferent
gonad is converted to a
testes.
• In the absence of SRY, a
different set of genes is
activated, and the
indifferent gonad becomes
an ovary.
• The germ cells, which
actually become sperm or
eggs, migrate into the
gonad about this time.
 Development of Phenotypic Sex
• The cells of the newly formed testes start secreting the
hormone testosterone.
– Testosterone secretion peaks about week 16, with levels similar to
those found in adult males. After this, the testosterone level
drops to about the same level as female fetuses.
– The testes also secrete another hormone: Mullerian inhibiting
substance (MIS) (aka anti-Mullerian hormone, AMH).
• Another important process in the developing male: during
the last trimester of pre-natal life, the testes migrate
(“descend”) from the kidney region into the scrotum.
– Under the control of a third testes hormone: “insulin-like
hormone 3”
• The developing ovary secretes estrogen, which is
important after birth, but estrogen from the mother
completely swamps it out before birth.
• In the early embryo, two duct Internal Ducts
systems form. After the gonad
differentiates into a testis or ovary,
one set of ducts develops further
while the other set degenerates.
• Testosterone causes the Wolffian
ducts to develop into male
structures: epididymus, vas
deferens, seminal vesicles.
– In the absence of testosterone,
the Wolffian ducts disappear
(except a bit becomes the adrenal
glands in both sexes)
• Mullerian inhibiting substance
causes the Mulerian ducts to
disappear.
– In the absence of MIS, the
Mullerian ducts develop into
the Fallopian tubes, uterus,
and upper vagina.
 Development of the External Genitalia
• This process is controlled by the presence
or absence of dihydrotestosterone (DHT).
• Testosterone gets converted into DHT by the
enzyme 5-alpha reductase, which is found in the
testes and the skin.
• Both sexes start out with the same structures,
which develop along different lines under the
influence of testosterone and DHT.
• The default condition in female: in the absence of
DHT, the external genital structures develop along
female lines.

• DHT also causes hair loss: male pattern baldness.

Testosterone is converted to DHT locally. Rogaine
works by blocking 5-alpha reductase
• In the absence of DHT, the
External Development
genital swellings form the
labia majora; the genital
folds remain unfused and
form the labia minora; the
genital tubercle forms the
clitoris and the urogenital
sinus forms the lower part
of the vagina.
• With DHT present, the
genital swellings migrate
and become the scrotum;
the urogenital folds
enlarge and enclose the
penile urethra and become
the shaft of the penis; the
genital tubercle becomes
the glans penis; and the
urogenital sinus forms the
prostate gland
 Childhood and Puberty
• During childhood, sex hormone levels are very low in both sexes.
– There is a surge of sex hormones in both boys and girls for a few weeks just after
birth. Significance is unknown.
• Puberty begins when the brain and hypothalamus start producing the
neurohormone GnRH (gonadotropin releasing hormone). This
hormone then stimulates production of LH and FSH by the pituitary
gland.
• LH and FSH stimulate the testes and ovaries to start producing large
amounts of testosterone and estradiol (a form of estrogen).
– In boys, some of the testosterone is converted to estradiol, which causes a growth
spurt, and sometimes leads to temporary breast development.
• The adrenal glands also secrete male sex hormones, in both boys and
girls, starting in late childhood.
– After puberty starts, the ovaries also produce androgens.
 Variant Conditions

• The large majority of people develop as either completely male

or completely female. However, 1% or more of the population
has some variant condition.
– Chromosomal variations
– Gene mutations
– External conditions
 Chromosomal Variants
• Meiosis, the form of cell division that generates the
sperm and eggs, carefully puts exactly 1 copy of each
chromosome pair into each cell.
• Sometimes meiosis goes wrong and puts 0 or 2
copies of some chromosome into a sperm or egg cell.
– the best example of this: Down syndrome, which starts
with a sperm or egg with 2 copies of chromosome 21.
– Maternal age effect: more frequent in older mothers
• The sex chromosomes are quite tolerant of variants.
• Most common types involve 45 or 47 chromosomes
• There are many other, rarer types, with 48 or even 49
chromosomes, such as 49,XXXXY. Such conditions
almost always lead to serious mental deficiencies.
• The general rule: if the Y is present, the person is
internally and externally male.
 Klinefelter Syndrome: 47,XXY
• Occurs about 1 per 500 male births. It is the most
common type of sex chromosome variant.
• The presence of the Y chromosome causes a 47,XXY
person to be male, both externally and internally,
because the testes are formed.
• Root symptom: small testes, leading to low
testosterone levels. Most, but not all, are sterile.
• At puberty, reduced facial and body hair, broader
hips, breast development.
• 47,XXY children tend to be taller, less physically
strong and coordinated, and more quiet and shyer
than their peers. Some language and learning
problems are common: often slow to learn to speak
and read.
• Testosterone replacement therapy helps with some 46,XX males, with the SRY
of the physical symptoms. Speech therapy and gene on the X, have the
educational services also help. Klinefelter appearance.
 Turner Syndrome: 45,X
• Only one X chromosome, sometimes called XO. Since
there is no Y chromosome, the primary gonad is the
ovary, and 45,X people are female.
• About 1 in 2500 live female births.
– 10% of all spontaneous abortions (miscarriages) are
due to Turner syndrome; about 98% of all Turner’s
embryos die before birth
• Ovaries completely non-functional, so 45,X women
are sterile, with no production of sex hormones and
development of secondary sexual characteristics at
puberty.
• Some characteristic physical abnormalities: short
stature, low hairline, webbed skin at neck. Kidney
and circulatory system problems
• Often have problems with spatial reasoning and You need 2 X chromosomes for
mathematics. Also social difficulties: inability to proper ovarian development.
understand others’ emotions. 46,XY females (non-functional
SRYgene) resemble Turner’s
• Can be treated with growth hormone and estrogen.
 47,XYY
• About 1 in 1000 live male births. Most XYY’s are never
detected: a very mild condition.
• since 1960, newly discovered chromosome variants aren’t given
the discoverer’s name
• It was once thought to create hyper-aggressive males with a
tendency towards criminal behavior.
– Richard Speck, the killer of eight student nurses in 1966,
pretended (falsely) to be an XYY to obtain leniency.
– A 1968 letter to the Lancet claimed that XYY men were in
prison at a rate "25-60 times as high as the prevalence in
the general population”, based on finding 2 XYY’s. 1970’s British TV series:
He had an extra Y, which
– the plot of Aliens 3 involves a prison planet for XYY’s.
made him a macho criminal!
• XYY’s are generally normal in appearance, but with average
height about 7 cm above expected and normal build. Perhaps
acne is more common than average, but this is disputed.
• They are often more physically active, somewhat delayed in
emotional maturity, and have a slight increase in learning and
speech problems.
• Fertile, normal sex drive, very rarely pass 2 Y’s to sons.
 47,XXX
• About 1 in 1000 live female births. So mild
as to be only rarely detected. Also called
triplo-X.
• Originally called “superfemale” (early
1960’s).

• Widely varying symptoms, including none at

all.
• Slightly more passive and quiet as babies,
less assertive, delayed motor and linguistic
skills. Delayed emotional maturity and social
skills. Some have slightly decreased
intelligence and learning difficulties.
• Lower back problems are common. Fertility
normal, don’t generally pass 2 X’s to
children.
• Both terms refer to people who have 2 different chromosome Mosaics and Chimeras
sets in different cells. For example, a 46,XX/47,XXY person has
some cells with 46 chromosomes and other cells with 47.
• A mosaic starts out with a single fertilized egg. During an early
cell division in the embryo, one cell gained or lost a
chromosome. (This is non-disjunction, the same event that
happens in meiosis to generate Klinefelter’s, etc.)
• A chimera starts out with two separate fertilized eggs, fraternal
twins. The two embryos fuse together to form a single
individual.
– It is not uncommon to have fraternal twins sharing some
blood cells, a “blood chimera”
– fused embryo chimeras are very rare: there are about 30-
40 known XX/XY chimeras (and undoubtedly an equal
number same sex chimeras). “tetragametic chimera”
– Chimerism is probably the way most true hermaphrodites,
who have both ovarian and testicular tissue, are formed.
However, actual XX/XY chimeras have been everything
from normal male, through various degrees of ambiguous
genitalia, to normal female.
• Sexual development can be quite variable in such people,
because the characteristics depend on which cells have which
chromosome complement.
 Gene Mutations
• The variants up to now all involve whole chromosomes, which have lots of
genes on them. The effects of changing the dosage of many genes tend to be
widespread but mild. (or completely lethal, as with most non-sex
chromosomes).
• Now we are going to look at several gene mutations. In these cases, only one
gene is affected, but it is completely knocked out. This can lead to large
effects, but limited to a few subsystems in the body.

• Rates are different: for chromosome changes, about 1 in 1000 births is a

typical frequency. For gene mutations, each parent needs to contribute a
mutated copy of the gene, so rates are usually 1 in 10,000 births or less.
• Inheritance is also a factor here: most chromosomal variants are spontaneous
events and don’t run in families. Gene mutations are usually inherited
variants: there is often a family/community history of the variant type.
– New mutations do occur spontaneously, but it’s rare. Most gene variants are
inherited from the parents.
 5-alpha Reductase Deficiency (5-ARD)
• 5-alpha reductase is the enzyme that converts testosterone into
DHT. If both copies of the gene that makes this enzyme are
defective, the person has 5-ARD.
– Recall that DHT is responsible for the development of male
external genitalia
• At birth, people with 5-ARD have undescended testes and male
ducts (with no female ducts), but genitalia that appear
somewhere between female and ambiguous, including a a very
small penis with hypospadias (which appears to be an enlarged
clitoris), and a short vagina. Often raised as girls
• At puberty, the increase in testosterone is large enough that
some DHT gets made, and they develop a male appearance: the
testes descend, the penis enlarges, facial hair appears, the voice
deepens, muscles develop.
• Large group in the Dominican Republic: maybe 1 in 90 men.
Called Guevedoces, a corruption of “huevos a los doce” (eggs--
testicles- at age 12). Raised as girls, they easily switch to the
male role.
– Other groups found in Malta, Jordan, Pakistan, New Guinea
 Androgen Insensitivity
• Incidence about 1 in 20,000 births
• Used to be called “testicular feminization”. 46,XY with normal
(undescended) testes. The testes secrete testosterone, but the cells
lack a receptor for it. No receptor = no response to the hormone.
Complete androgen insensitivity, CAIS.
• As a result, the male ducts (vas deferens, epididymus, seminal vesicles)
are not present. However, the testes secrete MIS, which causes the
female ducts (uterus, fallopian tubes, upper vagina) to degenerate .

• External genitalia develop as male if DHT is present, but

testosterone and DHT use the same receptor. So, female
external genitalia, including the lower 2/3 of the vagina.
• At puberty, the testes again secrete testosterone. The enzyme
aromatase converts it into estradiol. Thus, female secondary sexual
characteristics develop. Often “voluptuously feminine”. No
menstruation of course: no ovaries and no uterus. Pubic and armpit
hair is usually scant or absent.
– Occasionally, the undescended testes can become cancerous, so they are
often surgically removed after puberty is complete (so as to get normal
female development).
 Partial Androgen Insensitivity

• Sometimes, the testosterone receptors work

inefficiently, due to less drastic mutations than in CAIS.
In these cases, the body cells respond in a variable
manner to testosterone, leading a a wide variety of
ambiguous genitalia. PAIS = partial androgen
insensitivity. Also called Reifenstein syndrome.
– there is also mild androgen insensitivity (MAIS), which
leads to completely male appearance internally and
externally, but with some impairment of masculinization
at puberty.
• Variable symptoms: can be predominantly male (with
hypospadia, abnormal scrotum, small penis),
predominantly female (with enlarged clitoris, fused
labia, separate vaginal and urethral openings), or
ambiguous genitalia (microphallus--less than 1 cm
long), labia-like scrotum, hypospadia, gynecomastia.
• Similar variability in male internal ducts; females ducts
are usually absent due to MIS secretion.
• Sometimes people with PAIS change gender identity
after puberty, in either direction.
 Congenital Adrenal Hyperplasia
• The adrenal glands sit on top of the kidneys and secrete a variety
of steroid hormones, including cortisone (stress response),
aldosterone (salt balance) and androgens (male sex hormones).
• Steroid hormones are made from cholesterol through a series of
biochemical steps. Any one of these steps can be inactivated by
mutation. However, about 95% of CAH cases involve defects in
the enzyme 21-hydroxylase.
• 21-hydroxylase is needed to make cortisol and aldosterone (but
not androgens). Cortisol is secreted in response to the pituitary
hormone ACTH, in a feedback loop. So, if there isn’t enough
cortisol being made, more ACTH is made, and this causes the
adrenal gland to grow larger (hyperplasia).
• And, all of those steroid molecules that were destined to become
cortisol and aldosterone get diverted into male sex hormones
(androstendione and testosterone), which don’t need the 21-
hydroxylase.
• Very little effect on male fetus, which is already making
testosterone, except that after birth the lack of salt regulation can
lead to death from excess salt secretion (salt-wasting).
• Female fetuses with 21-hydroxlase deficiency have some
problems due to the flood of androgens released by the
adrenal gland. The ovaries are normal, and the female
(Mullerian) ducts are also normal (since no MIS is made). CAH
• Main effects are on the external genitalia: enlarged clitoris,
sometimes with an enclosed urethra (i.e. like the penis),
labia can fuse and become scrotum-like, vaginal opening
can be partly or completely closed.
• Appearance at birth varies a lot. Some appear to be normal
male with undescended (because non-existent) testes.
However, the chromosomes are XX, the gonads are ovaries,
and the uterus and fallopian tubes are usually intact.
• Normally, very little androgen is made in childhood. CAH
causes excess androgens throughout life, leading to rapid
growth, but an early closure of the bone growth plates: a
CAH is the most frequent cause
very short adult. Also: early puberty, with menstrual
of non-standard genitals in
problems (and poor sperm production in males).
genetically female (XX) children.
• The other hormones, aldosterone and cortisol, need to be
replaced. The cortisol replacement calms the ACTH activity,
leading to less androgen production.
 Some Environmental Causes
• Progestin-induced virilization. Progestin was used to prevent miscarriages in the
1950’s and 60’s. Related to this is the use of androgens to treat endometriosis
during that time period, and occasional accidental use of androgens. 160 known
cases.
– XX fetuses develop as normal females with functioning ovaries, but they may develop
some male secondary characteristics and often have enlarged clitorises. Effects are very
similar to CAH.

• Freemartin: usually seen in cattle: female and male twins, with testosterone from
male leaking over to the female due to a shared placenta. Normal female
appearance, but undeveloped ovaries and masculinized behavior. Rare or
unknown in humans.
– Aldous Huxley’s book Brave New World has human freemartins created by
hormone treatment of fetuses.
 And Lots More…
• We have covered the main causes of variation in
human biological sex. However, there are many
other, rarer conditions that also affect this.
• As always with biology, there are exceptions to
every rule, exceptions to everything I said here
today.
• What any individual feels about their body, and
how society reacts to these variations, is more a
matter for psychology and sociology than for
biology.
 Dosage compensation
• Ensures an equal expression of genes from the
sex chromosomes even though females have 2
X chromosomes and males have only 1
• In each female cell, 1 X chromosome is
inactivated and is highly condensed into a Barr
body
• Females heterozygous for genes on the X
chromosome are genetic mosaics

47
51
 Xist gene

X Chromosome
Inactivation

Barr body Heterochromatin formed,

genes silenced

http://embryology.med.unsw.edu.au/embryology/images/thumb/3/3f/X_inactiv
ation_Xist.jpg/400px-X_inactivation_Xist.jpg
 Process
• X inactivation center (XIC)
Near centromere
Contains 12 genes
 7 genes code for proteins
 5 genes code for untranslated RNA
(Rougeulle et al., 2003)

cen

XIC
 Process
• Xist and Tsix
Two genes actively involved in
inactivating an X chromosome
Antagonistic roles
 Xi   Xist expression
 Tsix expression
 Xa   Xist expression
 Tsix expression
(Lee et al., 1999)
 Process
• Xist and Tsix
Only genes expressed on Xi
X chromosome lacking Xist gene
cannot be inactivated

cen

Xist/Tsix in XIC
 Process
• Xist gene (pronounced “exist”)
Encodes a large RNA molecule
 Coats Xi from the XIC near
the centromere outward along the
X chromosome
(Lyon, 2003)

cen Xa

Xist RNA

cen Xi
 Process
• Mechanism for compacting Xi (Barr body)
Enzymes cause the following to occur:
 High levels of DNA methylation (CH3)
(Chadwick et al., 2003)

 Low levels of histone substitution of the

acetyl group (CH3CO) for a H atom in a
-OH group
(www.en.wikipedia.org/wiki/Acetylation)
 cen Xa

CH3 CH3 CH3 CH3

CH3
Xist RNA

cen Xi

CH3 CH3 CH3 CH3 CH3

 Process
High levels of histone H2A everywhere
(deNapoles et al., 2004)

High levels of histone H3 methylation

(Heard et al., 2001)

cen Xa

H2A

H3 cen Xi