FON 111 | EMBRYOLOGY NOTES

------------------------------------------------------------------------​by Sulaiman Al Salameh ‘​salsalmeh@alfaisal.edu​’

Introduction to
Embryology

● Embryology must be studied in a slow, systematic approach

● Earlier concepts must be understood clearly before moving on to more advanced concepts
● Embryology is studied to:
○ understand the root causes of human birth defects
○ understand the structural positioning of different organs
○ for early diagnosis and treatment
○ understand the molecular biology of human development (chromosome, stem cell, test-tube
baby, etc.)
● Although the cardiovascular system develops after the origin of the nervous system, it is the first
functional​ system developed in an embryo.

● Specific Objectives (FON 111 FINAL EXAM):

○ Describe gametogenesis
○ From the stage of fertilization, explain various stages of human development
■ During the first week
■ During the second week
○ Explain the development of chorionic sac, chorionic villi, amniotic cavity and yolk sac
○ Identify the etiology of birth defects (clinical embryology) in the first two weeks

● Meiotic cell division only occurs in the gametes

● Meiosis occurs in two stages and results in 4 haploid cells (n) with 23 number of chromosomes
● Diploid cells (2n) contain ​double​ the number of chromosomes = 46
● Four sperms form from one primary spermatocyte; one mature oocyte forms from the maturation of a
primary oocyte
○ the cytoplasm is conserved to form one large cell
○ polar bodies are nonfunctional cells that eventually degenerate
● !​ The ​second​ meiotic division of oocyte occurs ​after fertilisation

● Learn clinical terms related to the first week of development:

○ Nondisjunction​; disturbances in meiosis resulting in abnormal gametes
○ Anovulatory menstrual cycle
 ○ In vitro fertilisation (IVF)
○ Parthenogenesis; cleavage of an unfertilized oocyte
○ Surrogate mother
○ Abnormal embryos and spontaneous abortion
○ Common causes of infertility
● Understand the terminology!
○ read a textbook and learn the subject’s terminology

● Cleavage: series of synchronized mitotic cell divisions of the fertilized egg that results in the formation
of the blastomeres and changes the single-celled zygote into a multicellular embryo also
● The diagram on the left-hand side represents the first week of development; the diagram on the
right-hand side represents the. second-week of development

Gametogenesis-1
(Oogenesis)

Primordial germ cells (PGCs)

● From the second week​ of development onwards, the yolk sac accumulates primordial germ cells
which begin to be delivered to the developing gonads by the 4th week of development
● Until the ​10th week of development, the embryo freely floats​ in amniotic fluid and is connected to the
yolk sac
● The PGCs will migrate up from the yolk sac to the ​gonadal ridge; which will become later the sex
organs;
● Before the embryo decides which sex it is, it passes through the biopotential stage; if the embryo has
XX chromosomes, the​ gonads will differentiate into ovaries​ and the ​Wolffian duct will degenerate​, and
the​ Mullerian duct will differentiate into the fallopian tube (or oviduct)
● However, if a Y chromosome is present, ​the gonads will differentiate into the testes​, the ​Wolffian duct
will develop into the epididymis and vas deferens​, the ​Mullerian duct will degenerate
● Once the gonad starts differentiating into an ovary, the ​PGCs will differentiate into Oogonia​ (and
spermatogonia in males); these oogonia will undergo multiple rounds of mitosis

● Within the developing ovary, the surface epithelium of the ovary, forming the ​follicular cells, will start
to surround the oogonia​ as the oogonia attract them; as this happens, the ​primary oocytes will enter
the first round of meiosis
● Instead of proceeding to the 1st metaphase, ​primary oocytes are arrested at the diplotene stage
(resting period of prophase I)
● Follicular arrest is performed by a factor called: ​oocyte maturation inhibitor (OMI) which is secreted by
the follicular cells
● Primary oocytes stay in this condition until puberty
● Follicular cells surrounding groups of oogonia will start to surround individual oocytes, there is an
insufficient number of follicular cells to surround all oocytes, ​so oocytes with no follicular cell covering
them will die and degenerate
● The current theory is that the female has the pool of oocytes which she will need throughout her life
before birth, contradicting research to this was published in 2004 showing a possibility for stem cells
in the ovaries to differentiate into new oocytes post-birth
Folliculogenesis - Primary Follicle
● Follicular cells become cuboidal and stratified granulosa cells resting on a basement membrane
separating them from surrounding ovarian connective tissue that forms theca folliculi.
● Theca Folliculi promotes the growth of blood vessels in the theca interna​ (secretory cell layer) which
contains glandular cells and small blood vessels; it provides nutritive support for follicular
development.
● The ​theca externa is composed of dense connective tissue​ with larger blood vessels
● Granulosa Cells associate with Primary Oocyte and both secrete glycoproteins and begin to form the
Zona Pellucida

Folliculogenesis - ​Secondary Follicle

● Fluid-filled (antral) spaces​ appear between granulosa cells due to an osmotic gradient
● Joining of these spaces from the follicular antrum, which grows larger as the follicle matures further

Folliculogenesis - Graafian Follicle

● The granulosa cells surrounding the oocyte begin

forming the cumulus oophorus
● As the follicular antrum increases, the cumulus
oophorus decreases, leaving the oocyte
surrounded by a ​layer of Granulosa cells termed
the Corona Radiata

● Throughout maturation, the critical hormone

affecting growth is the FSH, another hormone is
LH
 ● For each ovarian cycle, ​15-20 primary oocytes undergo oogenesis;​ however, only 1 oocyte is ovulated
each cycle
● Just before ovulation, LH surge initiates completion of Metaphase I; ​LH works by inhibiting the arrest
factor OMI
● Meiosis I is completed and the second round of meiosis starts, also known as meiosis II

● Excess DNA following MI is transferred to the smaller polar body

● The Polar Body lies in the pre-vitelline space (between the oocyte and zona pellucida)
● The oocyte ​undergoes MII, but arrests at the second metaphase​ ~3 hours before ovulation
● Fertilization with sperm alleviates MII arrest; thus, technically, the female gamete only truly becomes
haploid (n) after fertilisation

Gametogenesis-2
(Spermatogenesis)

● Male Gonad: Testis

● Bipotential stage → gonads differentiate into the testis
● Mullerian duct degenerates
● Wolffian duct differentiates into the vas deferens (sperm is stored before ejaculation) and epididymis
(sperm sto​rage)
● Primordial germ cells differentiate into spermatogonia
● Spermatogenesis begins following onset of puberty; whereas oogenesis begins before birth and
completes after onset of puberty
● PGC differentiation is caused by a surge in testosterone

Testicular Organization
● Sperm is formed in the seminiferous tubules
○ At puberty, LH stimulates Leydig cells to produce testosterone, initiation spermatogenesis
○ Testosterone stimulates sertoli cells to provide nourishment for developing sperm cells, and
isolates sperm cells from the blood (immune cells in the blood will recognize haploid gamete
cells as foreign and destroy them; thus it is important to isolate developing sperm from the
blood)
○ Leydig cells = stroma cells; Sertoli cells = supporting cells

Spermatogonia
 ● Truly haploid cells come about only after
meiosis II which forms spermatids (truly
haploid cells)
Mature Sperm/ Spermatozoa
1. Acrosome formation (golgi apparatus is
involved in the formation of acrosome)
2. DNA condensation
3. Excess cytoplasm is shed
4. Flagellum formation

Motility:
● Sperm move to the epididymis where it is
stored until ejaculation
● They undergo surface and tail modifications
to develop the ability to fertilise
● Surface modifications in the epididymis:
○ Dropping of cytoplasm that was shed
○ More proteins are deposited on the sperm head for protection
Semen Production
● Sperm & seminal fluid: both are derived from seminal vesicle and prostate gland
● Seminal vesicles produce an alkaline solution consisting of fructose, fibrinogen, vitamin C, and
prostaglandins
● The prostate gland produces approximately half of the seminal fluid, consisting of citric acid, proteolytic
enzymes (help liquify coagulated sperm after it has been deposited in the vagina)
Capacitation:
● Sperm surface is modified inside female reproductive tract (in vagina) in order to better recognise and
fuse with the oocyte
Hyperactivation:
● Occurs in the cervix; it results in sperm flagella beating more rapidly and vigorously
Abnormal gametes can cause infertility
● e.g. nondisjunction (disturbances in meiosis) can bring about genetic diseases such as down syndrome
for example
Comparison of oogenesis and spermatogenesis
● Sperm is produced in an uninterrupted sequence; oogenesis has a long pause
● Spermatogonia continually self replenish; oogonia have a limited supply
● One oogonia results in one daughter oocyte; one spermatogonium results in four daughter sperm cells
The First Week
of Development

● Fertilization should occur near the ampulla of the fallopian tube (~1/3rd of the way to the uterus)
● Fertilized zygote moves to the uterus via contractions and peristaltic movement; oocyte transported
along fallopian tube through motion of fibrae and cilia on the epithelium

Ejaculated sperm in semen

● Human ejaculate ranges from 2.5ml to 5ml
● There is around 150-300 million/ml
● Semen contains
○ many types of cells and debris;
proteins and nutrients such as factors;
■ decapacitation factors;
● Motility (ability to move) and morphology (size and shape) differentiate good and bad sperm
● Taxis (movement)
1. Muscle contractions​ cause the sperm from the vaginal canal to move into the uterus
2. Within the fallopian tube​ thermotaxis direct sperm to the egg by an increased temperature of the
oviduct relative to the uterus​ (thermal-induced movement)
3. Chemotaxis directs the sperm to the oocyte by releasing chemicals​ close to the oocyte
(chemical-induced movement)
● Biochemical maturation of the sperm occurs in the female reproductive tract
● Sperm acquire motility and ability to fertilize as it swims to the oocyte
● Hyperactivation is needed in order for the sperm to break through the tough coating of the oocyte:
○ Mature oocytes surrounded by a thick glycoprotein layer called the zona pellucida
○ Further coated by a layer of cumulus cells, held together by an extracellular matrix

● Hyperactivation and capacitation occurs as a results from an influx of calcium ions in the sperm cell
● CatSper is a channel embedded within the plasma membrane of the sperm; this is a ligand-gated ion
channel that is opened by the action of the hormone progesterone; the opening of CatSper causes and
influx of calcium ions which in turn causes the processes of hyperactivation and capacitation to occur
inside the female reproductive tract
● The acrosome is a vesicular compartment immediately beneath the plasma membrane; it is thought to
contain digestive enzymes such as hyaluronidase and acrosin
● The acrosome reaction involves the release of digestive enzymes which aid in the removal of the
cumulus cells and the digestion of the zona pellucida;
● Membrane remodelling exposes sperm/oocyte binding proteins
● The acrosome reaction enables penetration of sperm of the cumulus cells and the zona pellucida
● Ejaculation → capacitation → hyperactivation → acrosome reaction
● The sperm/oocyte fusion:
○ Sperm contacts the oocyte, causing the oocyte wraps around the whole sperm
○ The nucleus of the mature ovum is called the female pronucleus, which fuses with the male
pronucleus to form a zygote.
● Following fertilization, the secondary oocyte completes meiosis II (alleviation of meiosis II arrest),
forming a mature ovum and a polar body
● Cortical granules release in the space between the zona pellucida and the oocyte; eventually, forming
a barrier to prevent polyspermy
● Calcium causes oocyte activation; the fertilising sperm causes elevation in calcium ion concentration
● The sperm nucleus is remodelled to allow fusion with the female nucleus (decondensation process);
this processes is calcium-dependent
● Embryogenesis starts and equal cell divisions occurs
● Embryo compaction starts to occur to from a morula
● A blastocyst is a hollow ball with inner cell mass, and an outer trophoblast
● The blastocysts hatches/ bursts out of the zona pellucida
● The hatched blastocyst floats in the uterine environment for two days
● Implantation begins about 6-7 days after fertilisation
● The implantation window lasts for approx. 3 days
○ Surface of endometrial epithelial cells are modified (decidualization)
○ LH from corpus luteum causes formation of pinopodes on the endometrium
○ Trophoblast microvilli interact with pinpodes
● Trophoblasts differentiates further into syncytiotrophoblast which secretes
metalloproteases/collagenases, digesting endometrial cell extracellular matrix
● The syncytiotrophoblast plays the most active role in invading the endometrium of the mother’s uterus
● During the invasion, endometrial blood vessels and endometrial glands are eroded, and a lacunar
network is formed
● Cytotrophoblasts deliver nutrients to the inner cell mass
● Syncytiotrophoblastic cells eventually envelopes the entire embryo
● Implanting blastocyst trophoblast cells synthesise secrete Human Chorionic Gonadotropin hCG
● hCG binds to LH receptors on ovarian luteal cells, sustaining progesterone levels
● Infertility is a huge problem worldwide; research shows infertility is mostly attributed to the male side
The Second Week
of Development

Implantation
● The uterus is composed of the
perimetrium, myometrium, and
endometrium
● Two layers are identified within the endometrium:
○ (1) the functional (epithelium) layer, which is sloughed off at menstruation
○ (2) the basal (CT) layer, which is retained at menstruation and serves as the source of
regeneration of the functional layer
● During the pregestational phase of the menstrual cycle, the functional layer undergoes dramatic
changes;
○ Uterine glands enlarge, and vascularity increases in preparation for blastocyst implantation
● The blastocyst floats freely in the uterine secretions for about two days
● Implantation begins on about the sixth day after fertilization
● The blastocyst implants in the functional layer of the uterine endometrium
● The outer cell mass (trophoblast) starts dividing rapidly, and at the embryonic pole, differentiates into
two layers:
○ Inner layer of cytotrophoblast (still dividing)
○ Outer layer of syncytiotrophoblasts (multinucleated)
● Finger like processes of syncytiotrophoblast begin the implantation process
● The blastocysts is superficially implanted to the wall of the endometrium by the end of day 7
● During week 2 of development, the embryoblast receives its nutrients from endometrial blood vessels,
endometrial glands, and decidual cells via diffusion
● Diffusion of nutrients does not pose a problem, given the small size of the blastocyst during week 2.
● Although the beginnings of a uteroplacental circulation are established during week 2, no blood vessels
have yet formed in the extraembryonic mesoderm to carry nutrients directly to the embryoblast (this
occurs in week 3)
Day-8
● Blastocyst partially embedded
● Trophoblast has differentiated into cytotrophoblast & syncytiotrophoblast
● Mitotic figures in cytotrophoblast
● Embryoblast is differentiated into hypoblast & epiblast (Bilaminar germ disc)
● Amniotic cavity appears within epiblast
● Amniotic cavity is lined by amnioblasts & epiblast
● Endometrial stroma is edematous & highly vascular
○ The large, tortuous glands secrete abundant glycogen & mucous

Day-9
● Blastocyst more deeply embedded
● Penetration defect in the surface epithelium is
closed by a fibrin coagulum
● At the embryonic pole, vacuoles appear in the
syncytium, fuse & form large lacunae (Lacunar stage)
● Hauser's or exocoelomic membrane originates from hypoblast
● A new cavity called exocoelomic cavity or primitive yolk sac is formed & it replaces blastocele or
blastocystic cavity

Days 10-12
● Blastocyst is completely embedded in the endometrium & produces a slight protrusion into the lumen of
the uterus.
● Lacunae form intercommunicating system which is more evident at embryonic pole
● Syncytiotrophoblast penetrate deeper & erode maternal sinusoids (capillaries)
● Maternal blood enters the lacunae establishing uteroplacental circulation
● In the meantime, extraembryonic mesoderm appears between cytotrophoblast externally and amnion &
Hauser's or exocoelomic membrane internally.
● Origin of this mesoderm is probably from yolk sac cells, hypoblast & epiblast
● Cavities appear within this extraembryonic mesoderm, fuse and form a new cavity called
extraembryonic cavity or chorionic cavity.
● This cavity surrounds the primitive yolk & amniotic cavity except where the germ disc is connected to
trophoblast by the connecting stalk (future umbilical cord)
● The extraembryonic mesoderm is differentiated into
○ Extraembryonic somatic mesoderm &
○ Extraembryonic splanchnic mesoderm
● Decidua reaction (cells & intercellular spaces)
● Growth of bilaminar germ disc is relatively low compared with that of trophoblast during this stage
Day-13
● By day-13, the surface defect has usually healed
● Occasionally, bleeding occurs at the implantation site as a result of increased blood flow into the
lacunar spaces
● Trophoblast is characterized by primary villi

Summary of Second week of development

Bilaminar Germ Disc
1. Trophoblast differentiates into cyto- & syncytiotrophoblast
2. Embryoblast forms epiblast & hypoblast
3. Extraembryonic mesoderm splits into somatic & splanchnic layers
4. 2 cavities form; the amniotic and yolk sac cavities

● Syncytiotrophoblast secrete hCG (human chorionic gonadotropin) Which can be detected by

radioimmunoassays which serves as the basis for pregnancy testing the end of second week
● Clinical Correlation:​ Ectopic tubal pregnancy:
○ Ninety percent of ectopic implantations occur in the uterine tube
○ Ectopic tubal pregnancies result in rupture of the uterine tube and internal hemorrhage, which
presents a major threat to the woman’s life
○ The uterine tube and embryo must be surgically removed; the symptoms may sometimes be
confused with appendicitis.