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.
 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.

2-cell stage 4-cell stage

8-cell stage Morula

Figure 1. Starfish cleavage stages (Magnification 400X)

 Figure 2. Amphibian cleavage stages (Magnification x2.5k)

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

Figure 3. Starfish blastula (x400) Figure 4. Amphibian blastula (x400)

 Early Gastrula Early Gastrula

Middle Gastrula

Middle Gastrula

Late Gastrula

Late Gastrula

Figure 5. Starfish gastrula ( x400 )

Figure 6. Amphibian gastrula (x400)

II. QUESTION FOR RESEARCH.

1. From where are micromeres derived? Why are they smaller than macromeres?
a. Micromeres are formed by a vegetal-shift in the orientation of the mitotic
apparatus at the vegetal pole within the blastomeres at the 8-cell stage. (Goto &
Keller, 2020) They are smaller than the macromeres because the four vegetal
cells undergo a highly-unequal division which produces four micromeres that
are much smaller than their sisters the macromere. (von Dassow, 2010)

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?
 a. The layer involved in yolk digestion is the endoderm (Balinsky, 1975). A few
distinguishable characteristics between the three germ layers is that the
ectoderm gives rise to the nervous system and the epidermis. The mesoderm, on
the other hand, gives rise to the muscle cells and connective tissue in the body.
And lastly, the endoderm gives rise to the gut and many internal organs.
(Molnar & Gair, 2015)

3. In amphibian gastrulation, are there any evidences of cephalization? Why or why not?
a. No, because during amphibian gastrulation it gives rise to ectoderm which will
form the brain afterwards but not during the gastrulation.

4. Into what cavity does the blastopore open? The gastrular slit? What becomes of each?
a. The blastopore opens into gut cavity. The archenteron eventually becomes the
cavity of the digestive tract, blastopore becomes the anus, and the mouth
arises. The gastrular slit will become the amniotic cavity. The folds will rise up
and unite dorsally, enclosing the embryo in a double-lined chamber or amniotic
cavity. (Keeton, Sircus, Dworken, & Hightower, n.d.)

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

a. The eccentric position of blastocoel is because of the macromeres. Since the
horizontal cleavages positioned toward the animal hemisphere, the newly
forming blastocoelic cavity will appear in an eccentric position above the level of
the equator and slightly toward the grey crescent side of the cleaving egg. It will
remain in this position, beneath the animal hemisphere, until it is later displaced
by the development of other cavities. (Rugh, 1951)

6. Explain the changes in size and position of the frog blastocoel during gastrulation.
a. During gastrulation, the blastocoel is displaced to the side opposite the dorsal lip
of the blastopore as the new cells enter the embryo. The blastopore lip expands
ventrally and laterally as the processes of bottle cell formation and involution.
The blastopore that widened develops lateral lips and a ventral lip which
additional mesodermal and endodermal precursor cells pass. Then the
blastopore will form a ring around the large endodermal cells that remain
 exposed on the vegetal surface. The yolk plug, which is the remaining patch of
endoderm will eventually be digested. (Molnar & Gair, 2015)

Bibliography
Balinsky, B. I. (1975). An introduction to embryology (4th ed.). Retrieved September 2021, from
https://openlibrary.org/books/OL5055303M/An_introduction_to_embryology
Goto, T., & Keller, R. (2020). Development, Growth & Differentiation. (M. Taira, Ed.) Japanese Society
of Developmental Biologists. Retrieved September 21, 2021, from
https://onlinelibrary.wiley.com/journal/1440169x
Keeton, W. T., Sircus, W., Dworken, H. J., & Hightower, N. (n.d.). Human digestive system. Retrieved
from Britannica: https://www.britannica.com/science/human-digestive-system
Molnar, C., & Gair, J. (2015). Concepts of Biology (1st ed.). BCcampus. Retrieved September 2021,
from https://opentextbc.ca/biology/chapter/13-2-development-and-organogenesis/
Rugh, R. (1951). The frog : its reproduction and development. New York : McGraw-Hill Book
Company, Inc. Retrieved September 2021, from
https://embryology.med.unsw.edu.au/embryology/index.php/Book__The_Fro
g_Its_Reproduction_and_Development_6 von Dassow, G. (2010, March 3).
Micromere formation. Retrieved from gvondassow:
http://www.gvondassow.com/Research_Site/Picture_of_the_week/Entries/2010/3/3_Micromere
_formation.html