Cleavage

• A series of mitotic divisions whereby the enormous volume of egg

cytoplasm is divided into numerous smaller, nucleated cells
• Cleavage-stage cells are called blastomeres
• During cleavage, however, cytoplasmic volume does not increase.
This decrease in the cytoplasmic to nuclear volume ratio is crucial in
timing the activation of certain genes
• As the rate of cleavage decreases, the blastomeres become motile,
and nuclear genes begin to be transcribed. This stage is called the
mid-blastula transition.
 Fig 22.69, Molecular Biology of The Cell
•Single large egg cell rapidly subdivides by repeated mitosis into
many smaller cells: blastomeres, without any change in total mass

•The very high rate of DNA replication and mitosis during cleavage.
•After about 12 cycles of cleavage (7 hours), the cell division rate
slows down and widespread transcription of the embryo’s genome
begins. This event is called the mid-blastula transition

• Embryo acquire many properties:

• the "gap" stages (G1 and G,) are added to the cell cycle
• the synchronicity of cell division is lost, because different cells synthesize
different regulators of MPF.
• new mRNAs are transcribed
From fertilization to cleavage
• Fertilization activates protein synthesis, DNA synthesis, and the cell
cycle.
• One of the most important events in the transition from fertilization
to cleavage is the activation of mitosis-promoting factor, or MPF.
 Rapid mitosis occurs during the Cleavage stage

Fig 5.1, Developmental Biology

Blastomeres generally progress through a biphasic cell cycle consisting of

just two steps: M (mitosis) and S (DNA synthesis).
The MPF activity of early blastomeres is highest during M and undetectable
during S.
 Rapid mitosis occurs during the Cleavage stage

Fig 5.1, Developmental Biology

The shift between the M and S phases in blastomeres is driven solely by

the gain and loss of MPF activity.

What causes this cyclical activity of MPF?

 What causes this cyclical activity of MPF?
• Mitosis-promoting factor consists of two subunits.

• The larger subunit, cyclin B, displays the cyclical behavior that is key
to mitotic regulation, accumulating during S and being degraded
after the cells have reached M

• The small subunit of MPF, the cyclin dependent kinase: Without the
cycIin B subunit, however, the cyclin-dependent kinase subunit of
MPF will not function.
 Cyclin dependent kinase.
• This kinase activates mitosis by phosphorylating several target
proteins, including histones, the nuclear envelope lamin proteins, and
the regulatory subunit of cytoplasmic myosin.
• It is the actions of this small kinase subunit that bring about
chromatin condensation, nuclear envelope depolymerization, and the
organization of the mitotic spindle.
 Rapid mitosis occurs during the Cleavage stage

Fig: Cell cycles of somatic cells and early blastomeres.

(A)The biphasic cell cycle of early amphibian blastomeres has only two states, S and M.
Cyclin B synthesis allows progression to M (mitosis), while degradation of cyclin B
allows cells to pass into S (synthesis) phase.
 Rapid mitosis occurs during the Cleavage stage

(B) The complete cell cycle of a typical somatic cell. Mitosis (M) is followed by an interphase stage.
Interphase is subdivided into G1, S (synthesis), and G2 phases. Cells that are differentiating are usually
taken "out" of the cell cycle and are in an extended G1 phase called ‘G0’ The cyclins responsible for the
progression through the cell cycle and their respective kinases are shown at their point of cell cycle
regulation.
Classes of cyclins

Figure 17-16 Molecular Biology of the Cell (© Garland Science 2008)

Targets of Cdk are determined by cyclin
Cytoskeletal mechanisms of mitosis
• Cleavage is the result of two coordinated processes.
• The first of these is karyokinesis, the mitotic division of the
cell's nucleus. The mechanical agent of karyokinesis is the
mitotic spindle, with its microtubules composed of tubulin
(the same type of protein that makes up the sperm
flagellum).
• The second process is cytokinesis: the division of the cell.
The mechanical agent of cytokinesis is a contractile ring of
microfilaments made of actin (the same type of protein that
extends the sperm acrosomal process).
 Cytoskeletal mechanisms of mitosis
Process Mechanical agent Major protein Location Major disruptive
composition drug
Karyokinesis Mitotic spindle Tubulin Central cytoplasm Colchicine,
microtubules nocodazolea
Cytokinesis Contractile ring Actin microfilaments Cortical cytoplasm Cytochalasin B

Role of microtubules and microfilaments in cell

division. (A) Diagram of first-cleavage telophase in
a sea urchin egg. The chromosomes are being
drawn to the centrioles by microtubules, while
the cytoplasm is being pinched in by the
contraction of microfilaments. (B) Confocal
fluorescent image of an echinoderm embryo
undergoing first cleavage (early anaphase). The
microtubules are stained green, actin
microfilaments are stained red.
Patterns of cleavage

Fig 22.68, Molecular Biology of The Cell

•The pattern of embryonic cleavage is determined by two parameters:

(1) the amount and distribution of yolk protein within the cytoplasm,
(Yolk rich region is vegetal pole and opposite is animal)
(2) factors in the egg cytoplasm that influence the angle of the mitotic
spindle and the timing of its formation.
• Rotation of the egg cortex after the injection of sperm is the major cause for
asymmetric egg division
Patterns of cleavage
The yolk-rich pole is referred to as the
vegetal pole; the yolk concentration in the
animal pole is relatively low.
The zygote nucleus is frequently displaced
toward the animal pole.

The amount and distribution of yolk

determine where cleavage can occur and the
relative size of the blastomeres.

In general, yolk inhibits cleavage.

When one pole of the egg is relatively yolk-

free, cellular divisions occur there at a faster
rate than at the opposite pole.
 Patterns of cleavage

isolecithal (Greek, "equal yolk") => These

eggs have sparse, equally distributed yolk.
Ex: eggs of sea urchins, mammals, and
snails.

In these species, cleavage is holoblastic

(Greek holos, "complete") meaning that the
cleavage furrow extend through the entire
egg.
Patterns of cleavage
Zygotes containing large accumulations of
yolk undergo meroblastic cleavage, wherein
only a portion of the cytoplasm is cleaved.
The eggs of insects have their yolk in the
center (i.e., they are centrolecithal), and the
divisions of the cytoplasm occur only in the
rim of cytoplasm around the periphery of the
cell(i.e., superficial cleavage).
The eggs of birds and fishes have only one
small area of the egg that is free of yolk
(telolecithal eggs), and therefore, the cell
divisions occur only in this small disc of
cytoplasm, giving rise to the discoidal pattern
of cleavage
Cleavage in sea urchins
Cleavage in sea urchins
•radial holoblastic cleavage
•The first and second cleavages are
both meridional and are
perpendicular to each other (that is
to say, the cleavage furrows pass
through the animal and vegetal
poles)
•The third cleavage is equatorial,
perpendicular to the first two
cleavage planes, and separates the
animal and vegetal hemispheres
from each another
Cleavage in sea urchins
•The four cells of the animal tier
divide meridionally into eight
blastomeres, each with the same
volume. These eight cells are called
mesomeres (fourth cleavage)
•The vegetal tier, however,
undergoes an unequal equatorial
cleavage to produce four large
cells, the macromeres, and four
smaller micromeres at the vegetal
pole (fourth cleavage)
Cleavage in sea urchins
•As the 16-cell embryo cleaves, the eight
"animal" mesomeres divide equatorially
to produce two tiers, an1 and an2, one
staggered above the other. (fifth
cleavage)
•The macromeres divide meridionally,
forming a tier of eight cells below an2.
Somewhat later, the micromeres divide
unequally, producing a cluster of four
small micromeres at the tip of the
vegetal pole, beneath a tier of four large
micromeres. (fifth cleavage)
The small micromeres divide once more,
then cease dividing until the larval stage.
Cleavage in sea urchins
•At the sixth division, the animal
hemisphere cells divide
meridionally while the vegetal cells
divide equatorially; this pattern is
reversed in the seventh division.
•At that time, the embryo is a 120-
cell blastula, in which the cells form
a hollow sphere surrounding a
central cavity, or blastocoel. From
here on, the pattern of divisions
becomes less regular.
Solid ball to hollow ball (blastula)

Blastula Fig 22-71, Molecular Biology of The Cell

•Tight junctions between the blastomeres begin to create an epithelial

sheet that isolates the interior of the embryo from the external medium.
•The intercellular crevices inside the embryo enlarge to form a single
cavity called blastocoel
Blastula formation in sea urchins

• The blastula stage of sea urchin development begins at the 128-cell stage
• (A) Formation of a blastocoel as cell division continues. (B) Soon after the rapid divisions
of cleavage end, the previously rounded cells unite to form a true epithelium. The
fertilization envelope can still be seen. As cilia develop, the blastula rotates within that
envelope. (C) The vegetal plate thickens, while the animal hemisphere cells secrete
hatching enzyme and allow the blastula to hatch from the fertilization envelope.
Blastula formation in sea urchins
 Rapid and invariant cell cleavages last through the ninth or tenth
division, depending on the species. By this time, the fates of the cells
have become specified, and each cell becomes ciliated on the region of
the cell membrane farthest from the blastocoel.
 This ciliated blastula begins to rotate within the fertilization envelope.
 Soon afterward, differences are seen in the cells.
 The cells at the vegetal pole of the blastula begin to thicken, forming a
vegetal plate.
 The cells of the animal hemisphere synthesize and secrete a hatching
enzyme that digests the fertilization envelope. The embryo is now a free-
swimming hatched blastula.
Micrographs of cleavage in live
embryos of the sea urchin
Lytechinus variegatus, seen from the
side. (A) The 1-cell embryo (zygote).
The site of sperm entry is marked
with a black arrow; a white arrow
marks the vegetal pole. The
fertilization envelope surrounding
the embryo is clearly visible. (B) 2
-cell stage. (C) 8-cell stage. (D) 16-
cell stage. Micromeres have formed
at the vegetal pole. (E) 32-cell stage.
(F) The blastula has hatched from
the fertilization envelope. The future
vegetal plate (arrow) is beginning to
thicken.
Cell fate determination

By the 60-cell stage, most of the

embryonic cell fates are specified

Particular blastomeres
consistently produce the same
cell types in each embryo, but
these cells remain pluripotent
and can give rise to other cell
types if experimentally placed in a
different part of the embryo
Cell fate determination
•The animal half of the embryo
consistently gives rise to the ectoderm
- the larval skin and its neurons.
•The veg1 layer produces cells that can
enter into either the ectodermal or the
endodermal organs.
•The veg2 layer gives rise to cells that
can populate three different structures
the endoderm, the coelom (internal
mesodermal body wall), and the non-
skeletogenic mesenchyme (secondary
mesenchyme), which generates
pigment cells, immunocytes, and
muscle cells.
Cell fate determination
•The first tier of micromeres (the
large micromeres) produces the
skeletogenic mesenchyme (also
called primary mesenchyme),
which forms the larval skeleton.
•The second-tier micromeres (i.e.,
the small micromeres) play no role
in embryonic development but
appear to contribute cells to the
larval coelom.
Gastrulation
• Gastrulation is the process of highly coordinated cell and tissue
movements whereby the cells of the blastula are dramatically
rearranged
• During gastrulation, these cells are given new positions and new
neighbors, and the multilayered body plan of the organism is
established.
• The three germ layers outer ectoderm, inner endoderm, and
interstitial mesoderm are first produced during gastrulation.
Origin of three germline

The endoderm derives from the most vegetal blastomeres, the ectoderm from the
most animal, and the mesoderm from a middle set that contribute also to
endoderm and ectoderm.
The coloring in each picture is the more intense, the higher the proportion of cell
progeny that will contribute to the given germ layer
Fig 22-70, Molecular Biology of The Cell
Modes of gastrulation
• Invagination (or intrusion), the process by which the wall of a one-layered
embryo gradually turns inward and forms an internal layer.
• Epiboly (or overgrowth), by which relatively large cells rich in yolk are overgrown
by the small ones and find themselves inside, forming an internal layer.
• Ingression (or settlement), by which certain cells move to the interior of the
embryo and settle under the superficial layer; immigration may be unipolar
(settlement from one place) or multipolar (from various places).
• Delamination, by which the cells split transversely, converting the one-layered
wall of the embryo to a two-layered one.
• Involution, the inturning or inward movement of an expanding layer so that it
spreads over the internal surface of the remaining external cells
Types of cell movements during gastrulation
Cell movements in gastrulation
Gastrulation in Xenopus
Cell Specification and Axis Formation
• Cell fates are specified by cell-cell signaling or by the asymmetric
distribution of patterning molecules into particular cells
• These unevenly distributed molecules are usually transcription factors
that activate or repress the transcription of specific genes in those
cells that acquire them.
Axis formation
• Embryos must develop three
important axes as foundations
of the body:
• The anterior posterior axis
• The dorsal ventral axis
• The right left axis
Cell Specification and Axis Formation
Asymmetric distributions of patterning molecules begin during cleavage
and generally follow one of three mechanisms:
• (1) the molecules are bound to the egg cytoskeleton and are passively
acquired by the cells that obtain this cytoplasm;
• (2) the molecules are actively transported along the cytoskeleton to
one particular cell; Or
• (3) the molecules become associated with a specific centrosome and
follow that centrosome into one of the two mitotic sister cells
Once asymmetry has been established, one cell can specify a neighboring
cell by paracrine or juxtacrine interactions at the cell surface
Cell Specification and Axis Formation
• Embryos must develop three crucial axes that are the foundation of
the body: the anterior-posterior axis, the dorsal-ventral axis, and the
right-left axis.
• The anterior-posterior (or anteroposterior) axis is the line extending
from head to tail (or mouth to anus in those organisms that lack a
head and tail),
• The dorsal-ventral (dorsoventral) axis is the line extending from back
(dorsum) to belly (ventrum),
• The right-left axis is a line between the two lateral sides of the body.