Embryology: study of embryos

Embryos - early stage; does not resemble the adult stage Gametogenesis egg and sperm
• Traditional: study of structural changes in embryonic development Fertilization
• Contemporary: study of embryonic developmental process of integrated complex phenomena Blastulation Cleavage
Gastrulation Histogenesis; pattern
Foundation for: formation
• development of new techniques for prenatal diagnosis and treatments Neurulation/organogenesis Differentiation
• therapeutic procedures to circumvent problems of infertility
• the mechanisms to prevent birth defects Morphogenesis Fetal growth
*improvement in prenatal development = improved long-term postnatal effects
Fundamental questions addressed in Devtl Biology:
Embryology to Developmental Biology: analysis of biological development • How does the fertilized egg give rise to an adult?
Answers: • How does that adult provide yet another body?
• Simple  complex General problems of Developmental Biology:
• Single cell  multicellular organism Problem of differentiation:
• Single cell  specialized cell (in function and structure) How does the same genetic information result to different cell types?
Major functions of development: How can the fertilized egg generate different cell types?
• Growth: generation of cell number Problem of morphogenesis
• Differentiation: cellular diversity within generation How do cells form ordered structures? positioning, timing?
• Morphogenesis: cellular order within generation How form and pattern emerge?
• Ensures continuity of life from one generation to the next *morphogenesis investigates how the regulation of cell fates contributes to the form and structure of
Phylogeny: evolutionary history the organism and its component parts
Ontogeny: gamete formation, development, post-hatching, larval, and adult periods Problem of growth (cell division)
How is cell division and growth regulated?
Modes of development: Problem of reproduction
Ontogenic development How are reproductive cells set apart during embryonic development?
• Mosaic development depends upon specific morphogenetic determinants in the one-celled zygote *only the germ cells pass characteristics on to the offspring
that are divided asymmetrically among the daughter cells Problem of evolution
*cell fate is determined by the components of the cytoplasm found in each blastomere How to changes is development create new body forms and what changes are possible?
*if part of the embryo is removed certain cell types would be lacking in later stage of development Why is the distinction between analogous and homologous structures important?
• Regulative development depends upon interactions between parts of the developing embryo The question of environmental integration
resulting in formation of different tissues; able to form an individual How is the organism’s phenotype influenced by the environment?
*signals: cell-cell interaction in development
Other concepts in embryology:
Major processes of development Guidelines: directive influences on development
• Cleavage division • Preformed guidelines
• Pattern formation (body axes) -present right at the start of ontogeny
• Morphogenesis -maternal effect genes
• Cell differentiation • Progressively-formed guidelines
• Growth -appear gradually in every step of ontogeny; zygotic genes
*Maternal genes; maternal effect genes; oocyte cytoarchitecture: patterning - gastrulation Wolff: embryological devt. occurs thru
Fate: range of cell types that a particular embryonic cell can give rise to progressing growth and devt;
*Dependent on cell asymmetries, unequal cytoplasmic determinants, inductive information, morphogenesis (development of
morphogens structures from structureless yolk
Oct4 is req. for maturation of inner cell mass ICM material
Cdx2 is req. for maturation of trophoectoderm TE =EPIGENETIC CONCEPT; theory of epigenesis
Potency: ability of cell to follow a devt pathway • Carl Ernst von Baer: 1792-1876 descriptive embryology, VITAL FORCE; similarities between embryos
Totipotent; Pluripotent; Multipotent; Unipotent of vertebrates particularly amniotes
Determination: gradual commitment to a fate • FATHER OF EMBRYOLOGY
Embryonic induction: evocative influence of cells Von Baer’s Law:
*capacity & competence The more general features that are common to all members of a group of animals developed earlier
Regulation than the more special features which distinguish the various members of the group
*regulative devt: potential of a cell is much greater than what is indicated in its normal fate (if • Ernst Haeckel: 1834-1919 ONTOGENY RECAPITULATES PHYLOGENY
controlled exclusively by inductive signals) vs, w/ Johannes Muller: reinterpreted Von Baer’s Law in light of evolutionary theory
*mosaic devt: embryonic cells can develop only accdg to their early fate (if controlled exclusively by Biogenetic Law: ontogeny is a modified recapitulation of phylogeny
cytoplasmic determinants) individual development progresses through the adult stages of the organism’s ancestors
Inevitability • Christian Pander: existence of germ layers
• apoptosis (PCD) - controls cell number by deleting cells which fail to partner, also eliminates • Heinrich Rathke: 1820’s affinity between embryos of higher and lower vertebrates pharyngeal
dangerous and abnormal cells pouches
• biological clock - Oscillatory gene expression (notch signaling) somites; cell-intrinsic timers: change • Schleiden & Schwann: 1839 laid the foundation of Modern Embryology and Histology
precursor cells over time • August Weismann: germplasm theory, self-reproducing determinants as guiding force for
• presence of homeobox genes: masterdevelopmentalcontrol genes that act at the top of genetic morphogenesis, chromosomes
hierarchies regulating aspects of morphogenesis and cell differentiation in animals *the offspring inherits from GERM CELLS not the soma
Drosophillia HOM genes = mammalian Hox g. • Oscar & Richard Herwig: fertilization, polar bodies, advances in understanding of meiosis
• Intracellular clock • Hans Spemann & Gilde Mangold: induction; organizer effect (DLB)
• positional information
Differentiation MODELS
FROG Wilhelm Roux
Early History SEA URCHIN Hans Driesch
• Aristotle: 1st systematic study of embryos FRUITFLY Drosophila melanogaster Thomas Hunt Morgan
• William Harvey & Graaf: 1672 Ovarian follicle; Graafian follicle
• Hamm & Leuwenhoek: 1677 observed human sperm ART = Assisted Reproduction Technology
Renaissance History IVF Edward and Sleptoe 1978
• William Harvey: 1578-1657 omne vivum ex ovo promoting existence of ova ICSI - intracytoplasmic sperm injection 1990
• Antoine van Leewenhoek: 1632-1723 “animalcules” in semen; tiny preformed human; theory of
preformation Approaches in the study of Embryology
• Spalanzani & Wolff: 1733-1794 Descriptive Embryology
Spalanzani: both M and F sex products -Involves detailed study of structure and arrangement of minute internal organs
for initiation of devt; first artificial -Concerned with explanations of structural features
insemination (frog eggs) -Investigates when and how a process is carried out
Comparative Embryology Gametogenesis
Establishes relationship between developmental stages Series of changes where the Germplasm specialized generative cells  Primordial germ cells (PGCs)
Experimental Embryology  Gametes specialized sex cells
-Finds out why a process is carried out at a specific time in a specific manner
-Ascertains which activate or regulate devt process PHASES:
Chemical Embryology 1. Generation of germ cells and migration to the gonads
involves biochemical investigations of the embryo; ushered in molecular biology • Arise outside the gonads
Development Biology • Recognizable at early stage of development
Broader approach from embryonic development to postnatal development Anuran amphibians:
normal growth Unfertilized egg - germplasm
metamorphosis Cleavage: Vegetal pole
regeneration Gastrula: Endoderm (origin)
tissue repair Amniotes (birds, reptiles, mammals)
PGCs in the yolk sac endoderm
-large, high content of alkaline
phosphatase
Humans: recognizable 24d
Mouse: PGCs originally in epiblast of
gastrula (ectodermal in origin); localized
the region in ExEmbryo mesoderm;
*Development of PGCs depend on signals: Pluripotency markers Oct4+, NANOG, and SOX2
(responsible for ICM (Oct4+) and Trophoectoderm
induced by (BMP) Bone morphogenetic protein: signaling factor inducing formation, proliferation,
and development of PGCs proximal-posterior epiblast

1. PGC
2. Allantois
3. Rectum
4. Ectoderm
5. Foregut
6. Primordial heart
7. Secondary yolk sac
8. Endoderm (yellow)
9. Mesoderm (red)
10. Amniotic Cavity
PGCs Pathway (ridges: chemo attractants)
PGCs @ base of allantois
Yolk sac endoderm (PGCs become ExEm)
Newly-formed hindgut (via allantois)
Dorsal mesentery
Left and right genital ridges (Em proper)
*Teratoma- unidentified tissue, mitosis, cells not following the normal pathway

2. Multiplication of germ cells in the gonads (mitosis), proliferation of germ cells (mitosis)
• PGC to oogonia/spermatogonia
• Human F: mitosis stops early (before birth)
• @ birth - oocytes; ATRESIA: degeneration
• 400-500 oogonia
• Human M: mitosis is continuous

3. Reduction in the number of chromosomes by half (MEIOSIS)

Meiosis I: reductional division; primary oocyte (2n)
Prophase I
• Leptotene: chromonemata: threadlike chromosomes
• Synaptene/Zygotene: synapsis or pairing of homologous chromosomes
• Pachytene: pairing is completed; tetrad formation; crossing over
• Diplotene: chiasmata: point of crossing over: exchange of genetic material
• Diakinesis: new genetic components
Metaphase I: metaphase plate: random arrangement of HOMOLOGOUS CHROMOSOMES
Anaphase I: Homologous chromosomes separate (reduction from 2n to n)
Telophase I:
Meiosis II: equational division; secondary oocyte: “tids” (n)
Nondisjuntion
2nd mitotic division
Ex: chromosome 21 - down syndrome
Physical manifestation of nondisjunction
4. Maturation and differentiation
Spermeiogenesis
PGCs  spermatozoa
Via mitotic multiplication, meiosis, and spermeiogenesis
Mitosis occurs throughout life
PGCs at genital ridge becomes:
sex cords  Seminiferous tubules  tubular epithelium  Sertoli cells

Cytoplasmic bridges: join cells in each succeeding division until individual sperm are separated from
residual bodies
Categories of changes during spermeiogenesis:
1. Reorganization of the cytoplasm
 Golgi apparatus
 Centrioles
 Mitochondria
2. Progressive reduction in nuclear size
 Nuclear elongation
 Loss of water
 Elimination of RNA; leaving only DNA
*Sperm maturation among vertebrates that undergo external fertilization differ from those that
undergo internal fertilization

Sidelights &speculations

BMP8B (bone morphogenetic protein 8B)

 Synthesized by spermatogonia
 Regulates the initiation of spermatogenesis during puberty
Oogenesis
PGCs to Mature oocytes
Via Cellular, molecular, and physiological > phenomena

Distinct features of oogenesis

 1O oocyte undergo arrested stages
 Unequal cytoplasmic divisions during meiotic divisions
 Only one functional oocyte produced per cycles (polar bodies)

Mammalian oogenesis:
PGCs
Fetal period Oogonia Mitosis No Follicle
Before or at birth Primary Prophase I: LZPD - Primordial Follicle
After birth Oocytes (Meiotic Arrest I) Primary Follicle
Diakineses
Metaphase I
Anaphase I Secondary
After puberty Telophase I Follicle
Secondary Prophase II Tertiary Follicle/
Oocyte; Metaphase II - Graafian follicle
1st polar body (Meiotic arrest II) – corpus luteum
Anaphase II & corpus
Ovulation Telophase II albicans
Fertilization – Mature
completion of 2nd Oocyte;
meiotic division 2nd polar body
Control of Human Reproduction LH: stim development of ova, induction of ovulation, and stimulation of estradiol and progesterone
https://opentextbc.ca/biology/chapter/24-4-hormonal-control-of-human-reproduction/ production by the ovaries.
Hypothalamus: Controls release of hormones from the pituitary gland. Estradiol: secondary sex characteristics
• Sends a gonadotropin-releasing hormone (GnRH) to the anterior pituitary which releases Estradiol + progesterone: regulates the menstrual cycle
follicle stimulating hormone (FSH) and luteinizing hormone (LH) to blood.
Ovarian cycle:
Male Hormones: • Governs the preparation and maintenance of the uterine lining
At puberty: • 22-32 days average of 28 days
FSH and LH enters the testes and stimulates Sertoli cells to begin facilitating spermatogenesis
using negative feedback 1. Follicular Phase and proliferation phase
LH enters testes, stimulating the Leydig cells to make Slow rising of FSH and LH = growth of follicles on surface of ovary (preparing the egg for ovulation;
and release testosterone into testes and the blood follicles release estrogens and low level of progesterone (maintains the endometrium to help ensure
Testosterone = secondary sexual characteristics, stimulates pregnancy).
spermatogenesis. Deepening of voice, facial hair, axillary and pubic
hair, sex drive Stage of development: Morula
Inhibin: inhibits FSH and LH when sperm count is too high 30-60 cells

No pregnancy = lining sheds, estrogen levels rise, the endometrium begins to regrow, replacing the
blood vessels and glands that deteriorated during the end of the last cycle.

High level of estrogen causes FSH and LH to rise rapidly, then fall. The LH spike causes ovulation: the
most mature follicle ruptures and releases its egg. Other follicles are lost. Estrogen decreases.

2. Luteal Phase and Secretory phase

Follicle: cells inside forms corpus luteum (produces estrogen and progesterone)
Uterus: being prepared to accept a fertilized egg
Only 1 egg: inhibition of FSH and LH, progesterone is elevated
No fertilized egg: corpus luteum degenerates, estrogen and progesterone decrease. Endometrium
degenerates. Low progesterone, allows hypothalamus to send GnRH to anterior pituitary, releasing
FSH and LH initiating next menses

Female Hormones:
Release of FSH and LH from anterior pituitary
Release of estrogen and progesterone from developing follicles

Estrogen: reproductive hormone, secondary sexual characteristics; breast development, flaring of

hips, shorter period necessary for bone maturation
Progesterone: assists in endometrial re-growth and inhibition of FSH and LH release
FSH: stim development of ova, develops in follicles
Inhibin: produced by follicle, inhibiting FSH production
Avian:
Amphibian
Fertilization:
 Ex vivo
 In vivo
Insemination: deposition of sperm cells into the F reproductive tract 2. Regulation of sperm
1. Contact and recognition between sperm and ova

Cortical reaction

A. Sperm entering cervical mucus at external os of cervix

B. Sperm interacting with endosalpingeal epithelium of the fallopian tube
C. Hyperactivated motility of sperm in fallopian tube
D. Oocyte in ampulla of fallopian tube
Entry into the ova. 3. Fusion of genetic materials of M and F
Transport of gametes & fertlization in mammals:  Protamines
Fertilization site: Ampulla of fallopian tube  Amphimixis-pronuclei fusion
Sperm Transport in F reproductive tract:
 Site of insemination
 Barriers in reproductive tract
 Uterus
 Entrance into the uterine tube
 Final destination
 Capacitation
Union of gametes
In Mammals:
 Hyaluronicdase from acrosomal cap digests corona radiata ZP + PM
 Sperm-specific receptor present in ZP
*ZP3 proteins in rodents

1 2

3 4
4. Activation of M metabolism to start development

Accomplishment of fertilization
 Completion of the second meiotic block
 Restores normal diploid chromosomes
 Sex of the future embryo is determined
 Variation
 Metabolic activation of the egg

ICSI: Intracytoplasmic sperm injection

GIFT: sperm and eggs are placed in a fallopian tube to allow fertilization in the natural site. The
woman must have at least one normal, open fallopian tube.
 *MPF is periodically active because Cyclin B is broken down during mitosis and re-synthesized
Cleavage: during S phase
 Generates large number of cells Mitosis is dependent on availability of cyclin
 Generates many copies of the zygotic genome
 Segregates cytoplasmkic components into different blastomeres
 (maternal cytoplasmic factors=Cytoplasmic gradients
 Increases the nuclear:cytoplasmic volume ration
o Universal aspect of cleavage;
o Ratio is different per species
o At a certain level, rate of cell division slows down
*Synchronous
*Asynchronous : no growth, but increase in umber of cells (mammals)
**Zygote: fertilized oocyte

What fuels cleavage?

MPF (mitosis promoting factor
Translated from maternal mRNA
Cyclin B (large subunit)
Cdc2/cdk1 – cylin depended kinase (small subunit) Histone: can change configuration of DNA
Source of MPF in the early embryo?
How does MPF work? [phosphorylates]  Cyclin B is translated from maternal RNAs
All the other necessary components for cell cycling are also maternally derived
** the cell cycle is independent of the nuclear genome for several cell divisions
 Regulators of Cyclin B resides in the cytoplasm of the egg

Maternal to Zygotic genome transcription: To replenish depleted egg cytoplasm and maternally
loaded cyclin B and other cell factors.
Beginning of new phenomena:
1) addition of gap phases
2) Loss of synchronous division
3) New mRNAs are transcribed
4) New cells synthesize different regulators

When does the embryo stops dividing?

• A new balance in the nuclear:cytoplasmic volume ratio (efficiency)
• Depletion in the maternal reserve of mRNAs and proteins
Result to:
• Zygotic genome activation (ZGA) new gene transcription in the zygote nucleus
Synthesis and degradation of MPF = cycling of cells • Synchronous cell division is lost (becomes asynchronous)
Activation/deactivation of cdk
Cell cycle of a typical mature somatic cell:
includes M (mitosis) phase, S phase (DNA synthesis) two gap phases, G1 and G2 2. Amount and distribution of yolk
(when the cells grow, produce mRNAs and proteins necessary for cell cycle) Types of egg as to yolk content:
Cell cycle of early cleavage divisions:  Isolecithal/oligolecithal
gap phases do not occur  mesolecithal
the cells cycle between M phase and S phase (biphasic)  telolecithal
* if G2 would occur, it is short  centrolecithal
*** there is no growth

In normal cell cycles:

presence of the cyclin-cdc2 complex promotes the transition from G2 to M phase

In early cleavages:
the cyclin-cdc2 complex promotes the transition from S to M phase no Gap phase (period of growth)

Factors influencing Cleavage Pattern

1. Maternal cytoplasmic factors
Angle of mitotic spindle  Site of 1st cleavage furrow
 Invariably, cytokinesis occurs in a plane perpendicular to the axis of mitotic spindle
 Maternal gene products may orient mitotic spindles
Holoblastic Rotational
Avian cleavage to blastula

At the end of Cleavage: Bastula, blastula, blastocyst

Developmental Properties of Cleaving embryos

 Regulative development
 Totipotent
Gastrulation: “gaster” **Spatiotemporal gene expression: Needs to be expressed at specific time and position
1. Lay down precursor of primitive gut; Devt. Landmark: initial formation of blastopore
2. Onset of morphogenetic movement and cell rearrangement Formation of Germ Layers
Regulation:
Major movements:  Notochord and floor plate of neural tube  Shh & noggin  sclerotome  PAX1:controls
1. Invagination: localized deepening chondrogenesis and vertebrae formation
2. Epiboly: cell-division; spreading of cells  Dorsal neural tube  WNT proteins  PAX3 : restricts the dermamyotome
3. Involution: mass movement of cells inward through an os *WNT proteins direct dorsomedial portion of somite: differentiate into muscle cell precursors
4. Convergent extension: increase in one dimension  Muscle cell precursors  MYF5 (muscle specific gene)
5. Delamination: splitting of layers of cells  Dorsal neural tube  NT3 : middorsal portion of somite  dermis
6. Passive movement  WNT protein and BMP4 activates MyoD expression
Cell behaviors:
1. Migration
2. Intercalation
3. Ingression
4. Rate of cell division
5. Change cell adhesiveness
6. Apoptosis – PCD
3-germ layers:
 Ectoderm – epithelial tissue type
 Endoderm (closely packed together)
 Mesoderm - mesenchymal tissue (loose arrangement) extracellular matrix
**Epithelial-mesenchymal transition

Cell movement is governed by gene expression

1. Mitosis: MPF  proteins translated from gene exp
2. Morphoregulatory molecules:
a. Selector Genes:
i. CAMs : Cell Adhesion Molecules
Facilitates cell-cell contact designed for easy attachment and detachment
ii. SAMs : Substrate Adhesion Molecules
Facilitates adhesion of cells to extracellular matrix; contact guidance for
sensory migration
Morphoregulatory cycle:
A. controls expression of genes for CAMs and SAMs
B. Deployed when needed
C. Changes cell behavior
D. New cell-cell contact generates new signals (intracellular/intercellular)
E. Feedback to selector genes: repeats until body plan is formed even after gastrulation
Neurulation: Attached to notochord:
 Establish precursor of CNS MHP- median hinge point cells- remain attached to the notochord; doesn’t move
 Start organogenesis DHP- dorsolateral hinge point- becomes wedge-shaped
 Continuation of formation of posterior-anterior body axes
 Somites: mesoderm are migrating to the sides, coalesce to form the mesoderm Neural cells= migratory ex: ganglin – aggregates of neural cells

**Anencephaly: if ANTERIOR neuropore do not close: brain is exposed to amniotic fluid

**Spina Bifida: POSTERIOR neuropore doesn’t close BODY AXES FORMATION
1. Anterior-Posterior
Modes of neurulation ; depends on species 2. Dorsal-ventral
1o Three cells amphibians, mammals at tail end 3. Left-right
2o solid rod of cells, closely packed; avian mammals
PATTERN FORMATION:
Signaling factors of neurulation: Establishment of Spatial information: instructs the cell position relative to the body axis
BMP4[active] Starts gradually from gastrula
 Favors ventral development Ex: formation of the limbs
 Ventralization of mesoderm
o Lateral plate mesoderm Recall:
o Intermediate mesoderm – kidney level Neural tube: (varying concentrations=different level of exprosure to the signaling cells = differences
among cells) BMP4,7, dorsalin, activin
What deactivates? Ectoderm
Transformation growth factor B superfamily: Neural roof
BMP4[deac]  Exp in CM/HN (chordin, noggin, follistatin, dorsalin, activin)  goosecoid Neural floor – sonic hedgehog
gene (upstream regulation, (TF) binds to transcriptive active site of: (exp in CM/HN genes)
DV in amphibians
BMP4[inactive] Recall fertilization: V- yolk, AP: pigmented movement of pigment molecules towards the entry point
 Dorsal development: of sperm, creates GC region = future site of DLB;
o CNS, notochord from axial mesoderm = notochordal process
As early as fertilization: in amphibians DV is established
Prechordal cells: first group of migratory cells Signaling factor: B-catenin - expressed in dorsal side
Amnioblastic cells B-catenin+Tcf3  Siamoid and Twin gene > goosecoid gene @chordamesoderm  activates CM/HN
Trilaminar embryonic formation ecto meso (dorsal, lpm, intermediate) endo genes  act/deac BMP4
Oropharyngeal membrane: ecto +endo If active: ventralization intermediate/LPM
Cloacal membrane: ecto = endo If inactive: dorsalization = starts with thickening of notochord = notochordal process & dorsal
Gut: endo mesoderm
Spatiotemporal gene expression: must be tightly regulated at the right time/location
Extraembryonic : hypoblast if not = malformation
Placenta : trophoblast
Degradation/synthesis of B-catenine is dependent on GSK-3 if Brachyury misregulated= insufficient mesoderm, compromises organs mesodermal in origin
Phosphorylated: B-catenine PO4  enzymatic breakdown( Tcf3 can’t function)  deactivated siamoid (urogenital, limbs); caudal dysgenesis = can’t generate posterior part. could shorten embryonic disc
+ twin gene  downregulation of goosecoid  downregulation of chordin, noggin, follistatin 
dorsalization

Avian and Mammalian

Primitive streak: cranio-caudal/anterio-posterior establishment

Nodal gene expressed by Hensen’s node: establishes and maintains the primitive streak
When activated: it encodes for a transcription factor ( effect of transcription start site)
activates cascades of genes (will establish the cranial structures of embryo and other
signaling factors) OTX2 LIM1 HOX1  SF secreted by node Cerberus: suppresses
expression of nodal gene at cranial structures, restricts the genes, forming the
posterior end of the embryo

 HNF3 Hensen’s node factor 3 = establishing for regionality of the forebrain and midbrain
 FGF fibroblast growth factor = regionality of hindbrain
 Hox = further specifications ie other brain structures

Hensen’s node
+goosecoid: +chordin, noggin, follistatin  active BMP4  Ventral dorsal: LPM, IM
-goosecoid: -chordin, noggin, follistatin  deac BMP4  DH(?) notochordal process

Nodal cells = ciliated: beats, may favor concentration gradient on one side
Hensen’s node
5HT(Neurotransmitter; Serotonin) Nodal genes secrete  FGF8  triggers further expression of
Nodal gene and Lefty-2 genes (also in LPM) = gene products (proteins) will activate PITX2 = left-
sidedness +  activates Lefty-1 gene (expressed in roof plate of nt) = acts as barrier to prevent left-
sided signals from crosses to right and shh gene (expressed in floor plate of nt) = prevents expression
of left-sided genes on the right side

Mid-caudal region: Brachury T gene products formation of mesoderm towards the mid-caudal
region of the embryo;
Right-sidedness:
Candidate gene: snail gene and NXK3 gene expressed only on the right side
Goosecoid misregulation:
Overexpression:ventral
Underexpressed: neuaral tube defects, malformation of somites