Name: Desaliza, Chubby Mae D.

Section: BI5A

ABI 3110 – Developmental Biology Laboratory

Laboratory Exercise No. 3
Cleavage, Blastulation and Gastrulation

CLEAVAGE
Cleavage is the process characterized by the series of mitotic division performed by the
fertilized egg (zygote) that leads to the formation of a multicellular embryo. It is the onset of
developmental process wherein a unicellular gamete becomes a multicellular ball or disc called
blastoderm. Each cleaved cell is known as the blastomere.
There are several types of cleavage:
A) Holoblastic or Complete Cleavage
The egg cytoplasm divides entirely at different cleavage planes which may be meridional,
equatorial and latitudinal. This is classified into: Holoblastic Equal – The egg cytoplasm is divided
entirely by the first two cleavage planes meridionally at right angles followed by horizontal and
equatorial planes producing equal sizes of blastomeres. Examples: microlecithal eggs of amphioxus
and placental mammals. Holoblastic Unequal – The egg cytoplasm is divided entirely by the first two
meridional cleavage planes producing equal blastomeres up to 4-cell stage. Succeeding latitudinal and
meridional cleavage planes produce smaller blastomeres called micromeres and larger blastomeres
called macromeres. Example: mesolecithal eggs of amphibians
B) Meroblastic Cleavage or Incomplete Cleavage
Type of cleavage wherein the smaller caplike cytoplasmic portion of the egg divides partially
leaving a yolk laden area uncleaved. Examples: eggs of teleosts and elasmobranchs (superficial);
reptiles and birds (discoidal)

BLASTULATION
A process after the cleavage characterized by hollowing out of the blastoderm as influenced by
the internal arrangement of blastomeres forming a monodermic layer of cells surrounding the cavity:
blastocoel of segmentation cavity. The embryo formed is a hollow sphere of cleaved cells called
blastula
In Amphioxus, the differentiation of blastomeres into micromeres and macromeres are
pronounced in the 64-cell stage. From the vegetal pole to the animal pole, the successive layers of
blastomeres decrease in size. The blastocoel increases in volume and closes at both poles forming a
hollow cavity.
In Amphibians, a cavity is present at the center of the blastomeres. It increases in size as
cleavage progressed. It has a roof of small cells of the animal pole and larger yolk laden vegetal cells.
The roof of the blastocoel expands and cells become thinner. Cells from the center of the vegetal pole
moves upward until reaching the floor of the blastocoel.

GASTRULATION
This process is characterized by the rapid morphogenetic cell movements whereby the
presumptive ectodermal and endodermal cells of the blastula migrate to the interior. The embryo
elongates and make a 90 degree turn. The blastocoel is obliterated and a new cavity, archenteron is
formed which opens to the outside by means of a blastopore.

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 In Amphioxus, the invaginating layer of cells eliminate the blastocoel. A gastrula consists of an
outer layer of cells, the ectoderm and an inner layer, the endoderm and an intermediate layer, the
mesoderm. The cavity enclosed by the two layers is the archenteron (gastrocoel) which gives rise to
mesoderm and notochord dorsally and the entire alimentary tract ventrally. The opening leading to
the outside from the archenteron is the blastopore and its involution greatly help in the completion of
the gastrula forming a presumptive endoderm until it comes in contact with the ectoderm.

In Amphibians, three morphogenetic movements are involved: epiboly, involution and

invagination. These movements result in the obliteration of the blastocoel and formation of
archenteron. The cells associated with the archenteron are the chordamesoderm forming its roof and
endoderm forming its floor.
In placental mammals, gastrulation commences with the splitting of the inner cell mass of the
blastocyst into epiblast and hypoblast through the process of delamination. The endodermal layer
spread along the inner surface of the trophoblast forming the yolk sac surrounding the yolk that is not
there.

I. ILLUSTRATIONS.
A. Illustrate the cleavage stages of starfish and amphibians and label the parts.

Figure 1. Starfish cleavage stages (Magnification ______)

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 Cleavage of a frog egg. Cleavage furrows, designated by Roman numerals, are numbered in order of
appearance. (A, B) Because the vegetal yolk impedes cleavage, the second division begins in the animal region
of the egg before the first division has divided the vegetal cytoplasm. (C) The third division is displaced toward
the animal pole. (D-H) The vegetal hemisphere ultimately contains larger and fewer blastomeres than the
animal half. H represents a cross section through a midgastrula stage embryo.

Figure 2. Amphibian cleavage stages (Magnification ______)

B. Illustrate the blastula and gastrula stages of starfish and amphibian. Provide the magnification and
the appropriate section of your illustration. Label the parts.

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 Figure 3. Starfish blastula (______) Figure 4. Amphibian blastula (______)

Figure 5. Starfish blastula (______) Figure 6. Amphibian blastula (______)

II. QUESTION FOR RESEARCH.

1. From where are micromeres derived? Why are they smaller than macromeres?

Micromeres are transplanted from the vegetal pole of a 16-cell embryo into the animal pole of
a host 16-cell embryo. The transplanted micromeres invaginate into the blastocoel to create a new
set of primary mesenchyme cells, and they induce the animal cells next to them to become vegetal
plate endoderm cells. The transplanted micromeres differentiate into skeletal cables while the
induced animal cap cells form a secondary archenteron. Meanwhile, gastrulation proceeds normally
from the original vegetal plate of the host (Ransick & Davidson, 1993).
The small cells that form by complete unequal cleavage of an ovum. Micromeres differ from
the macromeres of the same embryo by their smaller size and yolk content. The micromeres are
usually found in the animal hemisphere of the embryo (for example, in frogs), although sometimes
they are located in the vegetal hemisphere (in sea urchins). First three divisions in urchin embryos
produce equal, or very nearly equal, daughters. So do the vast majority of subsequent divisions, but
at the fourth cleavage, the four vegetal cells undergo a highly-unequal division which produces four
micromeres that are much smaller than their sisters, the macromeres (Juliano et al., 2006).
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2. In the early stages of amphibian development, what layer is involved in yolk digestion? What is the
basis for the distinction between germ layers in gastrulation?
The amphibian egg, however, contains much more yolk. This yolk, which is concentrated in the
vegetal hemisphere, is an impediment to cleavage. Thus, the first division begins at the animal pole
and slowly extends down into the vegetal region (Hara, 1977). Germ layer, any of three primary cell
layers, formed in the earliest stages of embryonic development, consisting of the endoderm (inner
layer), the ectoderm (outer layer), and the mesoderm (middle layer). The germ layers form during the
process of gastrulation, when the hollow ball of cells that constitutes the blastula begins to
differentiate into more-specialized cells that become layered across the developing embryo. The germ
layers represent some of the first lineage-specific (multipotent) stem cells (e.g., cells destined to
contribute to specific types of tissue, such as muscle or blood) in embryonic development. Hence,
each germ layer eventually gives rise to certain tissue types in the body ( Winograd, 2020).

3. In amphibian gastrulation, are there any evidences of cephalization? Why or why not?

4. Into what cavity does the blastopore open? The gastrular slit? What becomes of each?

In the blastopore the gut cavity, or the archenteron eventually becomes the cavity of the
digestive tract, and the blastopore becomes the anus; the mouth arises as a new opening. The
primitive gut cavity, or archenteron, which eventually gives rise to the hollow core (lumen) of the
alimentary canal (Keeton, & Sircus, 2020). In the gastrular slit is the amniotic cavity. These folds,
consisting of extra-embryonic ectoderm and extra-embryonic mesoderm, rise up and fuse dorsally,
enclosing the embryo in a double-lined, fluid-filled chamber known as the amniotic cavity.

5. Explain the eccentric position of the early frog blastocoel.

The hollow cavity is called blastocoel which is eccentric in position due to macromeres and
hence also referred to as coeloblastula. The process of formation blastula is called blastulation.
Gastrulation: process of transformation of single layered blastula into a three layered gastrula. Since
the horizontal cleavages appear toward the animal hemisphere, this newly forming blastocoelic cavity
will appear in an eccentric position above the level of the equator, and slightly toward the gray
crescent side of the cleaving egg. It will remain in this position, beneath the animal hemisphere, until

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it is later displaced by the development of other cavities. The size of the blastocoel increases with the
formation of smaller and smaller surrounding cells ( Hill, 1951).

6. Explain the changes in size and position of the frog blastocoel during gastrulation.

During gastrulation, As the new cells enter the embryo, the blastocoel is displaced to the side
opposite the dorsal lip of the blastopore. Meanwhile, the blastopore lip expands laterally and
ventrally as the processes of bottle cell formation and involution continue about the blastopore. The
widening blastopore “crescent” develops lateral lips and finally a ventral lip over which additional
mesodermal and endodermal precursor cells pass. With the formation of the ventral lip, the
blastopore has formed a ring around the large endodermal cells that remain exposed on the vegetal
surface. This remaining patch of endoderm is called the yolk plug; it, too, is eventually internalized
(Balinsky, 1975).

Balinsky, B. I. 1975. Introduction to Embryology. 4th Ed. Saunders, Philadelphia.

Gilbert, S. (1970, January 01). Early Amphibian Development. Retrieved September 05, 2020, from
https://www.ncbi.nlm.nih.gov/books/NBK10113/

Hara K . The cleavage pattern of the axolotl egg studied by cinematography and cell counting. Wilhelm
Roux Arch. Entwicklungs- mech. Org. 1977;181:73–87. 

Hill, M.A. (1951). Embryology Book - The Frog Its Reproduction and Development 6. Retrieved from
https://embryology.med.unsw.edu.au/embryology/index.php/Book_-
_The_Frog_Its_Reproduction_and_Development_6

Keeton, W., & Sircus, W. (2020, February 04). Evolutionary development. Retrieved September 05,
2020, from https://www.britannica.com/science/human-digestive-system/Evolutionary-
development

Juliano CE, Voronina E, Stack C, Aldrich M, Cameron AR (2006) Germ line determinants are not
localized early in sea urchin development, but do accumulate in the small micromere lineage.

Mc Cauley, B. (2019). Animal Development. Retrieved September 05, 2020, from

https://brianmccauley.net/bio-6a/bio-6a-lab/animal-development

Ransick A , Davidson E H . A complete second gut induced by transplanted micromeres in the sea
urchin embryo. Science. 1993;259:1134–1138. [PubMed] [Reference list]

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Winograd, C. (2020, May 26). Germ layer. Retrieved September 05, 2020, from
https://www.britannica.com/science/germ-layer

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