DEVBIOL

LECTURE 4: CLEAVAGE TO BLASTULATION

Gliceria B. Ramos| 1st TERM | A.Y. 2022-2023
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OUTLINE
I. A IV. B
II.

One must show the greatest respect towards anything that

increases exponentially no matter how small…
Garrett Hardin (1968)
● Increases the ratio of nuclear volume: cytoplasmic
CLEAVAGE DIVISION volume
● Fertilization is the wake up call for the fertilized oocyte ○ Universal aspect of cleavage
to embark into the next phase of development -> ○ At a certain level, pace of cell division slows
Fast-paced action – cleavage division ■ *balanced volume or “setpoint”
● Species-specific
○ Cytoplasmic volume is gradually distributed
FUNCTIONS into smaller cells
● Generates large number of cells ○ Increase of nv, decrease of cv
○ Newly fertilized zygote (1 - 2 - 4 - 8 - 62…) ○ Species specific
until multicellular embryonic phase
○ Capable of moving relative to its other’s
position

● Generates many copies of the zygotic genome TRANSITION FROM FERTILIZATION (ZYGOTE) TO
○ Cleaving embryo - progeny of cells CLEAVAGE
○ Cell division undergoes mitotic division so ● Mitosis promoting factor (MPF)
each cell has their own copy of the genome ○ Fuels transition to cleavage
(genetically equal) ○ Translated from maternal mRNAs
■ Allows individual blastomeres the ○ Since oocyte has activated maternal mRNAs
freedom to express the subset of are no longer silenced
the genome ○ 2 subunits:
■ Guide in developmental pathway ■ Cyclin B
● larger
■ cdc2/cdk1 = cyclin dependent
kinase
● Smaller
● Homolog of cell division
control 2 (cdc2)

● Segregates cytoplasmic components into different

blastomeres (maternal cytoplasmic factors)
○ Not yet activated/fertilized - silenced
maternal mRNA ready to be translated into
proteins once activated by fertilization
○ Blastomeres acquire unequal/different
combinations of cytoplasmic components
■ Provide new cytoplasmic
environment to the cell’s nucleus
● Has influence on activity of
the cell - cytoplasmic
influence on nuclear
activity
● Can activate or deactivate
subsets of the gene → can
lead to developmental
pathway
HOW DOES MPF WORK?
● Subunits must be bound together as a complex
○ Only then is it active
● Periodically active
○ Used by mitosis then is degraded
○ Once degraded, it is synthesized again
○ Cyclin B is broken down during mitosis
■ Kinase is deactivated
○ Resynthesized during S phase
■ Then forms a complex with cdc2 to
be activated
● Cycling of cells
○ Cycle of synthesis and degradation of cyclin
B
○ Cycle of activation and deactivation of cd
kinases

1

● Kinase
○ Enzyme that phosphorylates a target protein:
■ Histone proteins → chromatin
condensation
■ Nuclear membrane proteins →
nuclear membrane depolarization
(break down)
■ Reg. cytoplasmic proteins →
spindle fiber organization (mitotic
spindle)
○ These result to the onset of mitosis →
cleavage
SOURCE OF MPF IN THE EARLY EMBRYO
● Cyclin B is translated from maternal RNAs
○ All the other necessary components for cell
cycling are also maternally derived (e.g.
cdc2, cdk1)
○ The cell cycle is independent of the nuclear
genome for several cell divisions
● Regulators of cyclin B reside in the cytoplasm of the
egg
​ ●
● Depletion of the maternally-loaded cyclin B and other
cell factors → zygotic transcription must begin
● Transition from maternal to zygotic genome
transcription
○ Beginning of new phenomena:
■ Addition of Gap phases
■ Loss of synchronous division
■ Transcription of new mRNAs
■ Synthesis of different regulators in
new cells
WHEN DOES THE EMBRYO STOPS DIVIDING
● Establishment of a new nuclear: cytoplasmic volume
ratio balance
● Depletion in maternal reserve mRNAs and proteins
This Results To:
● Zygotic genome activation
○ New gene transcription in the zygote nucleus
due to depleted maternal reserve
● Cell division becomes asynchronous
TIMING OF CLEAVAGE DIVISION
● Late embryonic cells has characteristics of mature
somatic cells
○ Presence of more growth phases
○ Slower; asynchronous
● Early embryonic cell cycle is biphasic
○ Only S & M phase is present at first
○ Very fast; no growth
○ Short G2 phase; synchronous
Cell Cycle of Typical Mature Somatic Cell:
● M phase, S phase, and Two gap phases (G1 and G2)
● When cells grow, they produce mRNAs and proteins
for the cell cycle
Cell Cycle of Early Cleavage Divisions:
● No gap phases
● Cell cycles between M and S phase only
CONTROLLER OF CELL CYCLE
● MPF - Mitosis Promoting Factor
● Must be in complex to be active
● In mature somatic cell, there are more regulators
(Cyclin A, B, D, E)
Has growth and gap phases
● 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
FACTORS INFLUENCING CLEAVAGE PATTERN
● Maternal Cytoplasmic Factors
○ Angle of Mitotic Spindle is believed to be the
site of 1st cleavage furrow
○ Invariably, cytokinesis occurs in a plane
perpendicular to the axis of mitotic spindle.
■ Contractile ring - for cytokinesis
○ Maternal gene products may orient the
position of mitotic spindles
● Amount and Distribution of Yolk
○ Types of egg as to yolk content:
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 ■ Isolecithal/Oligolecithal
● They contain small amount
of yolk, but evenly
distributed
■ Mesolecithal
● They contain moderate
amount of yolk, slightly
concentrated at one pole
■ Telolecithal
● There are some amount of
yolk that is highly
concentrated on one pole
■ Centrolecithal
● Concentration of yolk at
the center of the oocyte
■ Characteristic of vertebrates:
Isolecithal/Oligolecithal,
Mesolecithal, and Telolecithal
■ Characteristic of Aquatic and
Terrestrial Arthropods:
Centrolecithal
● Holoblastic radial
○ Proceeding from the animal hemisphere to
the vegetal, cleavage division slows down
■ Amphibian egg is slightly less
telolecithal
○ First and second blastomere are cut
vertically
○ Third blastomere = horizontal cut and more
displaced toward the animal pole
■ Displaced brought by the thick yolk
content
○ In the succeeding cleavage divisions,
another horizontal cleavage occurs that
cleaves the animal hemisphere faster than
the amphibian

PATTERNS OF CLEAVAGE & CLEAVAGE

SYMMETRY
● Holoblastic Cleavage
○ Isolecithal (avians) and Mesolecithal (frog)
○ Complete — proceeds from the animal
hemisphere going down to the vegetal
hemisphere
○ Types of Holoblastic Cleavage:
■ Radial
● Characteristic of
vertebrates
■ Rotational CLEAVAGE TO BLASTULA
● Characteristic of
vertebrates
■ Spiral
● Characteristic of
invertebrates
■ Bilateral
● Characteristic of
invertebrates
● Meroblastic Cleavage
○ Telolecithal
○ Incomplete — cleavage involves only the
active cytoplasm or blastodisc AMPHIBIAN CLEAVAGE TO BLASTULA
○ Types of Meroblastic Cleavage:
■ Discoidal ● Presence of gray crescent in the fertilized oocyte
● Characteristic of ○ Formation of gray crescent opposite the
vertebrates entry point of the sperm
■ Superficial/Meroistic ○ Gray crescent region is brought by cortical
● Characteristic of reaction during fertilization → causes the
invertebrates and mass movement of pigment granules toward
aquatic/terrestrial the sperm entry leaving the area appearing
arthropods lighter in color
○ Intermediate shade between the vegetal
pole and animal pole

● Around at least 128 cells, the developing and cleaving

● Frog Egg
○ Classified as slightly telolecithal
mesolecithal
● Birds, Reptiles, and Fish Egg
○ Classified as telolecithal type of oocytes
embryo starts to form an internal cavity called
blastocoel
○ Embryo has now embarked into the blastula
stage
● Distinct transition from late cleavage to blastula called
morula stage (solid ball of cell)
○ Cells start to secrete and pump fluid where
the pressure created inside will push the
blastomeres to the side creating a cavity
(blastocoel)
● Formation of blastocoel characterizes the blastuli
stage
● Three distinct regions at the end of blastuli stage: (1)
or gray crescent region; (2) vegetal region (heavy yolk);
(3) pigmented animal region (pigmented granules)

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 ○ Nanog
○ Stat 3

MAMMALIAN CLEAVAGE TO BLASTULA

● Holoblastic spiral
○ Characteristic of vertebrates

● Holoblastic rotational
● In mammals:
○ Cleavage is extremely slow; (during its
journey down the oviduct)
■ Slow cleavage can be explained by
evolutionary adaptation of
mammals
● Embryo being secured in
the uterus
● Unlike when fertilization is
external, where cleaving
embryo is exposed to an
embryonic and
unpredictable/perilous AVIAN CLEAVAGE TO BLASTULA
environment = fast
cleavage for amphibians
■ Undergoes mitosis and synthesis
phase
■ Same as early embryonic cell of
amphibians
○ Unusually asynchronous
○ Does not always proceed regularly from
2→4→8 blastomeres
■ Cell AB divides first horizontally
■ As it moves to another stage, it is
only until then that cell CD is
starting to divide horizontally or
vertically
■ Short formation of 3-cell stage but
transient

Remember, the mature yellow yolk or yellow yolk is actually the

egg cell for avians

● Compaction (at 8-cell stage) occurs
○ Maximized areas of contact
● Polarization (around 16-cell stage)
○ Outside cells (9-14)
○ Inside cells (2-7)
● Cavitation (Blastocyst stage - 32-64 cell stage)
○ Formation of the blastocoel
○ Forms the ICM (inner cell mass)
○ Polarized cells outside form the trophoblast
Looking at the top view of the yellow yolk or the egg cell is a
blastodisc and this is already a fertilized oocyte or a zygote; it
is now activated so it will now undergo cleavage division.
First plane of cleavage involves only the blastodisc and is
vertical
Second plane of cleavage may not be synchronous, but what
is known is that it may cut the first plane of cleavage at a right
angle to the first one, but still involves the blastodisc
Third plane of cleavage may not be asynchronous

What’s distinct is that in the fourth plane of cleavage, it goes

circumferential, and as a result, it forms centrally located cells
and peripherally located cells; Circumferential type of cleavage
is followed by another one and another one and another one,
which results in radial expansion of the blastomere; there will
be a lot of blastomeres at this point and there is radial
expansion if these blastomeres; the blastodisc that has started
cleaving and expanding radially, now forms the blastoderm;
● Pluripotency markers for the ICM once the blastdisc has started cleaving, it is now a distinct
○ Oct 4 blastoderm (Top view)

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At side view or sagittal point of view (cut sagittally), what you
will see in the side will be the formation of a subgerminal cavity

How is the subgerminal cavity formed? = the blastoderm will

now start to absorb the liquid or fluid that was once within the
subgerminal cavity, which makes it a cavity; what happens
after this is that the cells that remain on top start to fall off and
those that fall off will form another layer, which is the hypoblast,
and those left on top will form the epiblast

Epiblast are those left on the surface

Those that have fallen off and have formed another layer would
be called the hypoblast At the end of cleavage in avians, stacks of cell layers form at
the top
When a hypoblast splits or delaminates from the epiblast, a
cavity is formed. This is called the blastocoel, which is formed Area pellucida contains the blastoderm, and it is surrounded by
at the expense of the germinal cavity; basically, the germinal the area opaca, which is attached to the yolk
cavity splits and forms another layer in the subgerminal cavity,
so blastocoel is a cavity formed at the expense of the
subgerminal cavity AT THE END OF CLEAVAGE: BLASTOCYST

By the time the blastoderm starts splitting to form the epiblast

and hypoblast, this is already the signal of the transition from
the blastula to gastrula; there’s a swift transition from blastula
to gastrula; there’s a very thin line that separates the blastula
and early gastrula

Because the cells will fall off, the uppermost region will thin out,
and there’s a cavity underneath… when viewed under a
microscope, this region will appear light or translucent and it
becomes known as the area pellucida BUT there are also cells
that are still in close contact (sticking) with the underlying yolk
and when viewed under the microscope, they appear dark, this
area that appears dark is still enclosed abut with the underlying
yolk, and it is called the area opaca (opaque)

Area pellucida: translucent or lucid; it contains the epiblast, so At the end of blastocyst in mammalians, there are two distinct
this is where the embryo will form populations of cells:

Inner cell mass

Trophoblast

There is also formation of a blastocoel

AT THE END OF CLEAVAGE: BLASTULA

AT THE END OF CLEAVAGE: BLASTULA

Blastula: ball of cells with a cavity, which is the blastocoel

Animal pole is a distinct region where the blastocoel is closest

to
Altogether, at the end of:
Marginal zone is the gray Crescent area or gray Crescent Amphibian blastula= ball of cells with the cavity called the
region blastocoel
Avians=stacks of cell layers with the formation of the
Vegetal pole is heavy with yolk subgerminal cavity (epiblast and hypoblast)
Mammalian= two distinct populations of cells (ICM and
trophoblast)

AT THE END OF CLEAVAGE: BLASTULA

Developmental properties of cleaving embryos:

1. capable of regulative development

- Development of the embryonic cells based on cell to cell
interaction
2. Cleaving embryos have totipotent cells (in the early
cleavage stages)
- Totipotent=cells have total potential