EMBRYOLOGY LAB 2-4.

1 OUTLINE
Ascaris uterus
EXERCISE 2
Fertilization of a Microlecithal egg
FERTILIZATION & MATURATION STAGE Unfertilized Egg
1. Sperm Penetration Stage
Primary oocyte
2. Maturation Stage
Fertilization First polar body formation
o is the direct interaction and fusion of two germinal cells (egg and Male pronucleus
spermatozoa) Female pronucleus
3. Second polar body formation
4. Pronuclear stage
5. Fusion stage

2.1 FERTILIZATION OF A MICROLECITHAL EGG

Specimen: Ascaris (uterus)

o Parasitic nematode in the intestines of some mammals
o Y-shaped reproductive tract; begins with a coiled and threadlike ovary & enlarges
at the posterior region as the uterus
o The uteri of the two branches unite to form the vagina, which ends in the genital
pore or vulva Unfertilized egg
o During copulation & insemination: sperm cells enter egg cells in the germinal tract o Extensive membranes observed in fertilized eggs are absent here
before its final stage of oogenesis is completed
1. Sperm penetration stage
o Well- developed reproductive system
o Separate sexes o Presence of sperm head
o Fertilization occurs at the end of the uterus nearest the oviduct inside the egg
Small, dense and
triangular bodies
o Cell is in 1 oocyte
stage

Primary oocytes
o Have vacuolated
cytoplasm,
inconspicuous nuclei & thin cell membranes

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 o After fertilization: become surrounded by a fertilization membrane and thick
outer shell
o Meiosis II can only continue after this has happened
o Sperm heads may also be seen between oocytes
3. Second Polar body formation
o When 2 oocyte extrudes one half of its longitudinal split chromosomes
o 2nd polar body looks exactly like the 1st
Two black streaks observe = inner streak
Only one streak observed; female pronucleus is already formed

4. Pronuclear stage

o male pronucleus appears as a dense body near the center of the egg
o Perivitelline space
between egg and fertilization membrane; filled with fluid
5. Fusion stage
o female and male pronuclei migrate towards each other
o produces diploid zygote nucleus
2. Maturation stage o array of microtubules can be observed near the centrioles of the male and
female nuclei
o First polar body formation terminates the process of fertilization
Diploid number of 4 (2 groups of 4 chromosomes can be observed in the
cell) *Early cleavage
Chromosomes appear as bead-like structures
- occur inside the uterus
Tetrads are found in the periphery of the primary oocyte
May also show the extruded first polar body on its surface
Black streak on fertilization membrane
Sperm head transforming into a pronuclei
Male Pronucleus
o nucleus of the sperm after the penetration of the ovum and enlarges
o product of 1st meiotic division

Female Pronucleus
o nucleus of the ovum is a product of 2nd meiotic division

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 o Eggs of: arthropods, ghag fishes, bony fishes, reptiles, birds
Isolecithal
EXERCISE 3 o Isos equal
EARLY DEVELOPMENT o Even distribution of yolk in the egg
o A microlecithal egg is isolecithal
Telolecithal
o Has most of its yolk concentrated on one pole vegetal pole
The development of an organism involves an increase in mass and an elaboration of
o Amphibians moderately telolecithal
structures. Both of these changes can be understood in terms of cell activity, like an increase in o Birds highly telolecithal
cell number (due to mitosis), enlargement of cells, migration of cells, as well as differentiation
Centrolecithal
and specialization of cells
o Arhtropods especially insects
EARLY DEVELOPMENTAL STAGES: the yolk is concentrated in the interior of the egg
o Cleavage Cytoplasm is distributed as a thin coat on the external surface
o Blastula
SPECIMENS:
o Neurula
Cleavage o Microlecithal egg = Starfish egg
o A mechanism for cloning the zygote genome generated at fertilization following o Megalecithal egg = Chicks egg
male & female pronuclear fusion o Medialecithal egg = Frogs egg
o Occurs right after fertilization
o It includes the zygote undergoing a series of mitotic cell division OUTLINE
o Leads to the formation of zygote and blastula Microlecithal: Medialecithal: Macrolecithal:
o In animals: (where cleavage (cell division) involves the whole egg) the blastula Starfish egg Frog egg Chick egg
usually consists of a hollow ball of cells 1.Cleavage: 1.Cleavage:
Gastrulation Unfertilized egg Animal pole
Fertilized egg (micromere)
o The first time many organisms begin to express the new embryonic genome that
Cleavage division Vegetal pole
was formed at fertilization
stage (macromere)
CLASSIFICATION OF EGGS 2. Blastula Cortical cytoplasm of
Blastula the egg
o Acdg. to the relative amount of yolk which they contain Blastocoel Fertilization
Blastoderm membrane
Microlecithal / Oligolecithal Fertilization 2. Blastula
o Micro small, lekithos yolk membrane Blastula
o Little yolk 3. Gastrula animal
o Eggs of: echinoderms, coelenterates, amphioxus, mammals (except monotremes) pole/micromere
vegetal
Medialecithal / Mesolecithal pole/macromere
o Medius middle Blastocoel
o Moderate amount of yolk Blastoderm
o Eggs of: annelid, mollusks, lamprey, amphibians Fertilization
membrane
Megalecithal/Macrolecithal
3. Gastrula
o Megos large

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 Observe the 2-cell, 4-cell, 8-cell, 16-cell and morula stages. Take
3.1 CLEAVAGE note the arrangement of blastomeres
Blastos sprout, meros - part
Cleavage cells / Blastomeres
o Cleavage is not simply multiplication of cells but rather it produces cells which, 3.1.2. MEDIALECITHAL EGG
eventually, become arranged distinctly
o Due to the predetermined pattern influenced by the amount of the yolk o Amphibians
among vertebrates exhibit medialecithal/mesolecithal egg
Yolk larger than of the starfish egg due to the larger quantity of yolk
o Very important factor in the cleavage formation because its presence may distinctly divided into a dark pigmented hemisphere (animal
inhibit cell division pole/macromere) and a lightly or unpigmented hemisphere
(vegetal pole/micromere)
The eggs of Microlecithal or Alecithal (no yolk) and its yolk is equally distributed the pigmented granules reside in the cortical cytoplasm of the egg
throughout the embryo will be completely divided into blastomeres of equal size within the plasma membrane and are more abundant in the animal
In Medialecithal, the yolk are pulled to its lower position (the vegetal pole) hemisphere
Macromere yolk is distributed throughout the cytoplasm of the egg but is much
o Pole with more yolk that will have bigger and fewer blastomeres more concentrated toward the vegetal pole of the egg
o Cells have not yet divided as often Hence, the yolk offers resistance to cleavage in the vegetal
Micromere hemisphere
o Pole with less yolk that will have many small blastomeres The amount of yolk is still sufficiently small to allow complete
cleavage of the cell
3.1.1 MICROLECITHAL EGG
Identify:
Unfertilized Egg
o Inactivated ovum of the Starfish 2-cell, 4-cell, 8-cell and 16-cell stage
exhibits prominent prematuration nucleus Nucleus lightly stained structure at the central portion of each of the cells
Fertilization membrane a loose, filmy, structure surrounding the zygote
Germinal vesicle with a distant nucleus
Possesses a vitelline membrane
Difficult to identify because of its close adherence to the 3.2 Blastula
egg.
Formed by the egg and is a primary membrane 3.2.1 MICROLECITHAL EGG
Fertilized Egg
Blastula
o After fertilization, the zygote shows an elevated vitelline membrane termed as
o Blastos + coilos = cavity
the fertilization membrane
o A hollow ball of cells resulting from successive cleavages
o Perivitelline space space formed due to the separation of fertilization
Blastocoel
membrane from the egg
o Germinal vesicle of the fertilized egg has broken down in preparation for the o Centrally located cavity
o Walls consist of an epithelial layer of cells
maturation process
o Not readily identifiable because of the cells at the back part of the section is
o Sperm head may appear as a densely staining body within the egg cortex
still visible
Cleavage division stage
o The organization of the blastomeres at the external surface of the embryo is
o Starfish egg exhibits holoblastic radial cleavage
prominent

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 Blastoderm Gastrula
o Layers of the blastoderm are arranged at the side of the embryo and enclosing
o embryo
the blastocoel
Fertilization membrane o occurs after invagination at vegetal pole
o surrounding the blastula o epiboly, involution, invagination (3 kinds of movements)
o considered as the secondary membrane because it is not secreted by the egg
o groupings of cells with same developmental potentialities occur here
the animal and vegetal pole will not be identified for this specimen because it is not easy to o formation of ectoderm, mesoderm, endoderm (3 germ layers
identify these in oligolecithal eggs since the cells are almost of the same size
o Microlecithal - in-pocketing of the blastula with the formation of a double-layered
3.2.2 MEDIALECITHAL EGG cup
Blastula o Mesolecithal - formation of dorsal blastoporal lip within the gray crescent area
o A hollow sphere resulting from the segmentation of the zygote
o Chick & placental mammals - primitive streak formation
Animal pole
o Region where numerous smaller cells are found
Vegetal pole
3.3.1 MICROLECITHAL EGG
o Region where fewer but bigger cells are found
Blastocoel After the formation of blastocoel, there is plenty of space to move some of the surface
o Eccentric located cavity cells to the inside to form new layers
o Will be obliterated in the future
Flattened at vegetal pole (initial migration & differentiation of cells)
Micromeres
Cells at the animal pole invaginate within the blastocoel - internal layers (mesoderm & endoderm) of the cells
Macromeres are formed
Cells at the vegetal pole
Blastoderm Ectoderm - cells at outer surface of gastrula
o Layers of the blastomeres arranged at the side of the embryo & enclosing the Gastrocoel or archenteron - cavity created by invagination ; future primitive gut
blastocoel
Blastopore - opening of gastrocoel at vegetal pole
Fertilization membrane
Thin membrane surrounding the blastula
Secondary membrane Identify the ff. structures:
Not secreted by the egg but the follicle cells in the ovary
Gastrocoel
Chorion + vitelline membrane (if it exists at all) makes up the fertilization
New cavity formed by invaginated cells; become the digestive tract of
membrane
embryo
Term used to refer to one of the fetal membranes of the amniotes Early gastrula short gastrocoel
Late gastrula gastriocoel is half the length of embryo; apex
seen as thin walled vesicle
3.3 Gastrula Vesicle
diverticula or lateral pouches at the apical region of gastrocoel
Gastrulation
become separated from gastrocoel and give rise to many mesodermal
o morphogenic movement and well-ordered rearrangement of cells structures
coelomic sacs

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 Enterocoel
Cavity formed from outpocketings of gastrocoel
Blastopore
opening of gastrocoel to the outside ; marks the posterior end of embryo
future anus
Mesenchyme
large, stellate cells mesodermal in origin.
scattered between ectoderm and gastrocoel
arise by budding off from outer walls of gastrocoel
Bipinnaria larva
larva of starfish formed at the end of gastrula
presence of cilia covering the ectoderm of embryo
cilia is the principal locomotory organ of larva
3.3.2 MEDIALECITHAL EGG
Gastrulation
o invagination of advancing germ ring cells and continues to involution
o Involution 3.3.3 MEGALECITHAL EGG
inward movement of cells so that it spreads out to the internal layer
o cells at margin of germ ring - change shape so the surface layer turns inward
and begins to migrate anteriorly across the roof of blastocoel
o invagination of cells create archenteron
o blastopore - mouth of in-pocketing
*Dorsal lip - upper margin of curved fold
o as the in-pocketing deepens, the opening becomes a crescent, a horseshoe,
then a complete circle
PARTS:
o epiboly continues as the germ ring advances over the exposed yolk cells.
o By the time the circle is complete, yolk can be seen within the confines of the
blastopore in an area known as yolk plug
o blastopore is now surrounded by dorsal, lateral and ventral lips
o involution first occurred at the dorsal lip near the animal pole
o expanding layer of cells which form the roof of archenteron becomes the
endoderm and chordamesoderm
o for some time, the floor of archenteron remains composed of large yolk cells
o cells on outer surface of embryo is ectoderm
o involution at ventral lip of the blastopore
o gradually the invaginated endoderm proliferates to enclose the yolk and
complete the archenteron
o mesoderm soon develops between ectoderm and endoderm
Gastrula
2 layered embryo resulting from ectodermal movements of the cell
Gastrocoel / archenteron , blastopore
o Same in description in microlecithal
Lip of the blastopore
o portion of ectoderm where involution takes place
o Dorsal lip - more defined where involution is more prominent
o Ventral lip - less defined lower margin marked by a deep cleft
Yolk plug
o portion of vegetal pole that protrudes and obstructs the blastoporal
opening
o endodermal in origin
Blastocoel
o Remnant may be seen
Fertilization membrane
o Thin membrane that envelops the gastrula
Gastrulation starts with the thickening of cells in the posterior portion of area pellucida
As this happens, th cells inside the blastodisc split and proliferate into the blastocoels
The cells that spitted off are the hypoblasts whle the remaining cells on the top are the
epiblasts
Epiboly & involution hold true also in megalecithal eggs but in the manner they are
accomplished is different is different from medialecithal eggs
Area opaca darker, outer, peripheral area
Area pellucida clearer, central area; separated from the yolk by segmentation cavity
Primitive streak thickened part of the blastoderm; brought by the convergence of the
surface layer to the midline
Primitive groove central furrow
Primitive ridges thickened margin on both sides of the primitive groove
Primitive plate thickening at the caudal end of the streak
Primitive pit most cranial portion of the primitive streak
Primitive knot AKA Hensens node, anterior to the primitive pit; where presumptive
notochordal cells migrate
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 b. Presumptive Epidermis
EXERCISE 4.1
o remainder of the ectoderm
ORGANOGENESIS: NEURULATION o forerunner of the outer layer of the integument
c. Notochord
o oval mass of cells directly beneath the neural plate
o origin: mesoderm
Neurulation
o temporary
o Processes involved in the formation of various organs
o provides skeletal support
After Gastrulation d. Mesoderm
o middle germ layer
o cells are sorted out o formed in the embryo during gastrulation
o lose individual potentialities & become subdivided into parts o found on each side of the notochord and dorsal to the archenteric roof
o the start of the differentiation of the embryo into the tissue and organ primordia
from which the organ system that constitute the body of the organism will 3 REGIONS:
develop Epimere (dorsal mesoderm)
- form somites
Stages of Neurulation:
Mesomere (intermediate mesoderm/nephrotome)
1. Neural Plate Stage - form excretory system
2. Neural Fold Stage Hypomere (lateral plate mesoderm)
3. Neural Tube Stage - delaminates into the inner splanchnic and the outer somatic
mesoderm
e. Endoderm
o large, yolk-ladden cells that surround the gastrocoel

1. Neural Plate Stage earliest

Parts:
a. Neural Plate
o -neuroectoderm: region of embryonic ectodermal cells that lie
directly above the notochord
o aka medullary plate

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 3. Neural Tube Stage
o neural folds have fused at the middorsal region forming the neural tube with a
cavity called the neurocoel
o prescence of cilia and pigment granules of the embryo in the epithelial lining

PARTS:

a. Neural Crest Cells

- found at the edges of the neural plate and lies above the neural tube
- migrate throughout the embryo to form: ganglia, sympathetic nervous system,
pigment cells, parts of the gill arches, etc.

b. Differentiated Mesoderm

3 REGIONS:
Epimere
- segmented mesodermal blocks located on either side of the
notochord and lying beneath the epidermal ectoderm
- will differentiate into:
2. Neural Fold Stage neural folds are prominently elevated o Sclerotome forms axial skeleton
o Myotome forms muscle
Parts:
o Dermatome forms dermal layer of skin
a. Neural Fold
- aka Somites
- portion of the neural plate that has a depression and a laterally elevated
margin Mesomere
b. Neural Groove - fate: kidneys
- depression formed in the midline - stalk-like connection between the epimere and hypomere
- gives rise to the neurocoel - anteriorly: forms pronephros
c. Hypochordal -posteriorly: forms mesenchyme which develops into
mesonephros and metanephros
- group of cells present between the notochord and the archenteric roof
- aka intermediate mesoderm, nephrotome, nephromere
- temporary
-difficult to observe
- aka Subnotochordal rod Hypomere
- region posterior to the mesomere
- has split into 2:
o Outer Somatic mesoderm
o Interior Splanchnic mesoderm
(cavity formed between is called coelom)

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