CLEAVAGE

The Main Process of Animal Development

1
 KEY CONCEPTS

Cleavage, a series of rapid cell divisions

without growth, provides cellular building
blocks for development

2
What happens after fertilization?

24 hrs
1 day

5 days

7 days

3
 What happens now ?
 Development of the zygote, the study of
which is known as embryology or
developmental biology
 The zygote undergoes a series of mitotic cell
divisions called cleavage
 The stages of development are: Fertilized
ovum (zygote)  2-cell stage  4-cell
stage  8-cell stage  Morula  Blastula
 Early Gastrula  Late Gastrula
4
Developmental Stages in
an Amphibian

5
 What is Development?
 Includes all stages in the life of an individual
 Here, we focus on the early stages : zygote to embryo

 What happens during this time?

1. Fusion of egg and sperm forms the zygote
2. The zygote undergoes cleavage; mitosis increases numbers
of cells
3. Differentiation occurs via cell determination
a. The cells undergo changes in gene expression and so
change with time/place in the embryo; this is called
differentiation
b. Determination is the process of commitment of a cell
line to a particular developmental pathway
c. Some cells do not change; these are stem cells
6
 Differentiation and Morphogenesis
 The ongoing differentiation results in the development
of form and structure (i.e. morphogenesis) in the
embryo
 Occurs via changes in
 Cell protein expression
 Signaling between cells
 Cell migration
 Interactions with the extracellular matrix
 Controlled death (apoptosis) of cells
7
 Major types of morphogenesis movements seen early embryos
Type of Movement Description Example

Archenteron formation in
Invagination Inpocketing of a sheet of cells
Amphioxus

Outpocketing of a sheet of
Evagination Exogastrulation
cells

Rolling around a corner of an

Cell movement through
expanding outer layer of cells
Involution the amphibians
and spreading over an
blastophore
internal surface

Spreading of outer cells

Epiboly Spreading of a cell sheet towards amphibians
blastopore

Sinking of individual cells from Primary mesenchyme

Ingression
a surface into an area formation in Sea Urchin

Migration of cells as single

Delamination individuals through their own Migration of neural crest
motility

8
 The Types of
morphogenesis
movements

9
Stages of Development
Most animals proceed through
these stages during embryonic
development:

1. Zygote
2. Early cleavage stages
 Morula
 Blastula
3. Gastrulation
 Gastrula
4. Neurulation
5. Morphogenesis/Organogenesis
10
Embryonic development
11
Organogenesis
Organogenesis is the
formation of the organs.

The layers are germ

layers; they have
specific fates in the
developing embryo.

12
So, the processes of development are:

 Cleavage – cells undergo division (2, 4, 8 etc) without

the embryo increasing in size

 Growth – cells undergo division as well as increase in

size

 Differentiation – when cells take on specific structure

and function (the nervous system is the first visible
system)

 Morphogenesis – the embryo begins to take shape as

cells migrate
13
 Cleavage
 During cleavage the zygote divides by mitosis, giving
rise to many cells
 The egg contributes the majority of the zygote
cytoplasm
 Both gametes contribute equal numbers of
chromosomes
 The zygote forms a two celled embryo, and continues
divisions to form a ball of 32 cells called the morula
 The morula continues divisions to form the hollow
blastula with up to several hundred cells
• The cavity of the blastula is the blastocoel

14
• Cleavage is a series of rapid cell divisions without cell
growth or gene expression which occurs in early
embryogenesis

• Early cleavage divisions in most embryos are reductive

• During cleavage, the cytoplasm is divided into smaller

and smaller cells, called blastomeres

• The total cellular volume of the embryo stays the same,

but the number of cells within the embryo increases

• Cleaving cells have a modified cell cycle, in which the

two gap phases, G1 and G2, are completely omitted

• The cells cycle rapidly between M and S phases

15
THE CELL CYCLE

16
• The pattern of cleavage is influenced by the amount of yolk
in the egg

1. In eggs with less yolk, cleavages are equal, and the

resulting blastomeres are of similar size

2. If the yolk is localized, such as in frog eggs, then

cleavages are unequal - the cells derived from the yolky
region (the vegetal pole) are larger than those derived
from the region without yolk (the animal pole)

3. In the very yolky eggs of fish and birds, the cleavage

furrow is slowed, or even blocked, by the presence of the
yolk. Complete divisions are restricted to the least yolky
region of the egg, and the embryo forms as a cap of cells
sitting on top of the yolk
17
• Cleavage results in a blastula, a ball of cells with a
central cavity called the blastocoel

• Below are two diagrams of blastulas. On the left is a sea

urchin blastula, and on the right is a frog blastula

18
 Patterns Of Cleavage
The pattern of cleavage is affected by:
- quantity and distribution of yolk present, and
- genes controlling the symmetry of cleavage
Yolk is a mixture of proteins, phospholipids, and
fats
Amount and distribution of the yolk vary among
different animal groups

19
 Patterns of cleavage

1. Bilateral : Vertical/meridional & horisontal

2. Radial
3. Spiral
4. Circumpherencial

20
 Spiral cleavage is typical of protostomes:
annelids and molluscs
 After first two divisions the plane of cleavage tilts
and diagonal to the polar axis

 Radial cleavage is typical of deuterosomes:

echinoderms & vertebrates
 First division is vertical; second division is at right
angles
 Third division is horizontal at right angles to the
first and forms an 8 cell embryo with 4 above and
4 cells below the last division plane

21
Spiral Cleavage in a segmented worm embryo

22
 Radialcleavage
 Deuterostomes
 All vertebrates

23
• Early cleavage patterns vary widely between different groups of animals,
based largely on the orientation of the division planes. The simplest
pattern is radial cleavage, in which successful division planes are at 90
degree angles relative to each other. This results in the blastomeres
aligned directly over or to the side of one another

• In spiral cleavage, the division planes are not at 90 degree angles,

resulting in blastomeres that are NOT aligned directly over or beside one
another

24
= equal

= anequal

25
26
• Reptile and Avian eggs
Large amount of yolk concentrated at one end of
the of the cell (vegetative pole)
 Vegetal pole (nutrition)
 Animal pole (more metabolically active)
 Animal and vegetal poles
will later determine
the anterior/posterior or
dorsoventral
end of the embryo

27
 The Amniotic Egg
 Mamalian eggs have very little
yolk
 Extraembryonic membranes
protect and nourish the embryo
 Terrestrial vertebrates have
four extraembryonic
membranes
 Develop from the germ layers,
but are not part of the embryo
and are lost at birth
1. Chorion
2. Amnion
3. Allantois
4. Yolk sac 28
 Review : types of eggs
1. Amount of yolk
• Microlecithal : little yolk (lancelets and
(oligolecithal) placental mammals)

• Mesolecithal : moderate amount of yolk

(lampreys and amphibians)
• Macrolecithal : large amount of yolk (most
(polylecithal) fishes, reptiles, birds,
monotremes)

2. Distribution of yolk
• Isolecithal : evenly distributed
• Telolecithal : concentrated towards vegetal pole
• Centrolecithal : central distributed 29
Types of
Cleavage

30
Types of Cleavage

31
 Types of Cleavage

a - when cleavage complete and egg is divided into blastomeres.

b - eg chick where cytoplasm located at animal pole and only this region cleaves
c - eg: insects and crustaceans d - eg: mammals e - eg: molluscs,
32
worms
Cleavage in 3 different
animal lineages

33
The process of cleavage and the structure of the blastula are
both related to the amount of yolk present

1. Microlecithal eggs:
• Holoblastic cleavage
• Cleavage is equal
• Blastula is a hollow ball of cells with a blastocoel

2. Mesolecithal eggs:
• Holoblastic cleavage
• Cleavage is unequal
• Blastocoel is displaced into animal hemisphere

34
3. Macrolecithal eggs: (teleosts, reptiles, birds)
• Meroblastic cleavage
• Blastoderm : forms at animal pole

4. Placental mammals
• Holoblastic cleavage
• Cleavage is equal
• Trophoblast : superficial layer of cells
surrounding the blastula (these structures
collectively referred to as the blastocyst)

35
 Cleavage patterns in relation to yolk content of eggs

Types of Types of Pattern of Representative Blastula

Blastula
eggs cleavage cleavage animal groups cavity
Radial,
Sphere : Large
Oligolecithal- bilateral, Amphioxus,
Holoblactic wall a central
isolecithaal spiral, mammals
single layer sphere
rotational
Sphere :
wall layered
Small
Mesolecithal Holoblastic Radial Amphibians and of
sphere
nonuniform
thickness
Flat space
Cell disk on
Most fishes, between
Telolecithal Meroblastic Discoidal surface on
reptiles, birds epiblast &
yolk
hypoblast
Yolk-filled
Centrolecithal Meroblastic Superficial insects none
cylinder
36
Cleavage in
Sea Stars

37
Cleavage in Amphioxus

38
 Cleavage in Frogs
Animal pole

Vegetal pole

39
Cleavage in
Humans

40
Early Cleavage Stages in Birds and Reptiles
 Bird and reptile eggs have much
yolk concentrated at the vegetal
pole of the egg
 Cell division takes place in the
blastodisc
 Division is meroblastic; cells do
not completely separate from each
other and remain attached,
at least initially, to the yolk mass
 In birds and some reptiles, the
blastodisc splits into the epiblast
(upper) and hypoblast (lower,
nearest the yolk), separated by the
blastocoel 41
Cleavage in a Bird Embryo

Blastodisc

yolk

42
blastodisc

2-3 mm

43
formation of blastocoel
44
Cleavage
in Birds

45
Blastulation in Chick & Mammals

46
 Blastulation 1
After mitosis has created several hundred to a thousand cells,
a ball called a blastula is formed.

47
30hr 48hr

4
3 days days

4.5-5 6
days days

48
 Gastrulation

 The blastula develops a hole in one end and

cells start to migrate into the hole; this forms
the gastrula
 The process is called gastrulation

 Gastrulation results in a three-layered embryo

 The pattern of gastrulation is different in
different in different species and is affected by
the amount of yolk

49
50
 Gastrulation 1
 During the 3rd week, the two-layered
embryonic disc becomes a three-layered
embryo
 The primary germ layers are ectoderm,
mesoderm, and endoderm
 Primitive streak – raised dorsal groove that
establishes the longitudinal axis of the
embryo

51
 Gastrulation 2
 As cells begin to migrate:
 The first cells that enter the groove form the
endoderm
 The cells that follow push laterally between
the cells forming the mesoderm
 The cells that remain on the embryo’s dorsal
surface form the ectoderm
 Notochord – rod of mesodermal cells that
serves as axial support
52
 1. Microlecithal eggs:
• Invagination
• Blastocoel mostly obliterated, a double-walled
cup is formed
• Gastrocoel :opens via blastopore
• Rotation : animal-vegetal axis becomes
horizontal; dorsal surface of embryo is
uppermost
• Notochord forms from dorsal wall of
archenteron
• Mesoderm--from dorsolateral outpocketings of
archenteron; somites.
• Enterocoely : cavities of outpocketings become
coelom.
• Remaining part of archenteron is endoderm.
see slide 32 53
2. Mesolecithal eggs
• Process of involution; epibole -- growth of one part over
another.
• Blastocoel obliterated and two tissue layers formed.
• Gastrocoel (archenteron) -- opens via blastopore.
• Chordamesoderm -- forms notochord at midline and
paired blocks of mesoderm (somites).
• Remaining part of innermost tissue is endoderm; comes
back together dorsally after the formation of the notochord
and mesoderm.

54
55
3. Macrolecithal eggs
• Process of delamination.
• Epiblast -- upper sheet.
• Hypoblast -- lower sheet.
• Mesoderm formation.
• Sometimes by delamination.
• Mesenchyme -- cells detach from existing tissue and
individually form mesodermal tissue.
• Primitive streak--most of mesoderm formed this way;
surface cells stream toward streak were they involute and
spread out between original two layers.

56
57
Primary Germ Layers

58
4. Mammals
• Early embryo called a blastocyst.
• Two cavities in inner cell mass:
• Upper lined with ectoderm--forms amnion.
• Lower lined with endoderm--forms yolk sac.
• Mesoderm forms by delamination--spreads out to line
trophoblast.
• Flattened inner cell mass now a blastoderm

59
60
61
 Primary Germ Layers
 Serve as primitive tissues from which all
body organs will derive
 Ectoderm – forms structures of the
nervous system and skin epidermis
 Endoderm – forms epithelial linings of the
digestive, respiratory, and urogenital
systems
 Mesoderm – forms all other tissues
 Endoderm and ectoderm are securely
joined and are considered epithelia 62
 Germ Layers

 Ifdifferentiation
occurs, there may be
up to three layers.
 Ectoderm – outer
layer
 Mesoderm – middle
layer
 Endoderm – inner
layer

63
Gastrulation in Sea Urchin
Mitotic cell divisions Multicellular stage

mesoderm, diploblastic, triploblastic, enterocoely,

schizocoely, fate of the blastopore 64
Gastrulation in Chick & Mammal

65
66
67
68
 Gastrulation
 The blastula further grows and forms a gastrula.
 In more advance animals, the differentiation of
tissues begins.

69
 Gastrulation

 Ectoderm
gives rise to
outer covering
and nervous
system
 Endoderm
gives rise to the
digestive tract
 Mesoderm
gives rise to 70
 The vegetal pole
invaginates and meets archenteron
the opposite wall,
obliterating the
blastocoel
 The archenteron is the
newly formed cavity
 The blastopore is the
opening of the blastopore
archenteron, and
becomes the anus in
deuterostomes
Gastrulation in Echinoderms
71
 Gastrulation in Birds
 In birds, the epiblast cells form the primitive
streak, with the primitive groove serving as the
functional equivalent of the blastopore
 Cells migrate centrally, and dive into the streak, to
turn under the outermost layer of cells and move
laterally and anteriorly inside the developing
embryo
 No archenteron is formed
 Hensen’s node is the site of cells that will form the
mesodermally-derived notochord
 Cells sink into the interior and move under the epiblast
to form an extension from the node

72
Gastrulation in Birds

73
 Credit: © Carolina Biological/Visuals Unlimited

Chicken embryo, 21 hours. 74

 Neurulation

 The nervous system is the first organ system to

develop
 Once the nervous system is in place, the rest
of the organs can develop: organogenesis
a) The notochord grows and induces overlying
ectoderm to form the neural plate

75
b) Cells of the neural plate fold to form the
neural groove and the surrounding neural
folds

76
c) The neural folds fuse, forming a hollow neural
tube
 The anterior portion forms the brain; the rest
forms the spinal cord

77
 The Neural Crest
d) The neural crest is a critical structure that guides
formation of several organ systems
 The neural crest forms on either side of the point of

fusion
 Neural crest cells migrate to other regions of the

embryo
 At their destination they differentiate into many

different kinds of cells

78
Fig. 52.11

79
80
Cleavage in the zebrafish embryo is limited to a small region on "top" of
the yolky mass

A. 2 cell stage
B. 4 cell stage
C. 8 cell stage
D. 16 cell stage
E. 32 cell stage
F. 64 cell stage

81
(a) Unfertilized egg (b) 2-cell stage (c) 4-cell stage

Nucleus

100 µm 50 µm 50 µm

82
(d) 16-cell stage (e) Blastula (f) Early gastrula

Blastocoel

Blastopore
Archenteron
50 µm 50 µm 50 µm

83
Fig. 50-3 (d-f), p. 1084
(g) Middle gastrula (h) Sea star larva
(i) Young sea star

Archenteron
Mouth

Anus

Blastopore Stomach

50 µm 50 µm 1 mm

84
Radial cleavage in the sea urchin results in a hollow sphere of cells

At the right are pictures of various cleavage stages in the sea urchin
L. variegatus.
A. 1-cell zygote. The fertilization envelope is visible as a large "halo"
around the embryo. The arrow points to the site of sperm penetration.
B. 2-cell. C. 8-cell. D. 16-cell. E. 32-cell. F. Hatched blastula (F is
courtesy Dr. Chuck Ettensohn, Carnegie-Mellon Univ.)
85
 Amniotic Animals

 Includes reptiles, birds and mammals

Placental Mammals

Reptiles and Birds

86
 Cleavage in human
Degenerating Inner cell mass
zona pellucida
Blastocyst cavity
Blastocyst
cavity Trophoblast
(a) Zygote (b) 4-cell stage (c) Morula (d) Early blastocyst (e) Implanting
(fertilized egg) 2 days 3 days 4 days blastocyst
6 days

Fertilization
(a)
(sperm meets (b)
egg) (c)
Uterine tube
(d)
Oocyte Ovary
(egg)

Ovulation Uterus (e)

Endometrium
Cavity of
uterus

8728.4
Figure
(a) Male and female pronuclei prior to fusion 50 µm
88
 50 µm
(b) 2-cell stage 89
 50 µm
(c) 8-cell stage 90
~ 3 days

50 µm
(d) Cleavage continues giving rise to a morula 91
92
Similarities in Development Within Groups

93
 Differences
in
Development
Between
Groups

94
95
Animal Development

96
Starfish development, unfertilized egg. 4 blastomeres.
2 blastomeres.

Starfish development,
16 blastomeres. 32 blastomeres.
nonmotile blastula.
morula

97
Starfish development, gastrula during invagination. Starfish development, mid-gastrula. LM X75.

98
Starfish, late bipinnaria. Starfish, young adult.
 Mollusks

Annelids

Arthropoda

Echinoderms

Uchordates

Mammals
placenta

Aves
egg

Reptiles

Amphibians
The amniotic

99
 The chorion and amnion enclose the embryo
 The chorion surrounds the entire embryo
• in birds and reptiles it functions in gas exchange
 The amnion encloses the embryo and forms an open
volume between the embryo & the amnion called the
amniotic cavity
• The amniotic cavity fills with amniotic fluid, which envelops
the embryo and cushions it
• The amniotic fluid can be sampled to test for developmental
abnormalities
 The allantois is an outgrowth of the gut
• In reptiles and birds, it stores nitrogenous wastes
 The yolk sac encloses the yolk in vertebrates
with yolk-rich eggs
 In humans, there is no yolk sac, but the yolk aids in
formation of red blood cells
100
 Mollusks

Annelids

Spiral cleavage
Arthropoda

Echinoderms

Uchordates

Mammals

Aves

Reptiles
Radial cleavage

Amphibians
101
Organogenesis
Organogenesis is the
formation of the organs.

The layers are germ

layers; they have
specific fates in the
developing embryo.

102
 Organogenesis is the formation
of the organs
 Endoderm
Organogenesis
• The innermost layer
• Goes on to form the gut
 Mesoderm

• In the middle
• Goes on to form the
muscles, circulatory
system, blood and many
different organs
 Ectoderm

• The outermost
• Goes on to form the skin
and nervous system

103
From mesoderm arise the Somites

 Blocks of mesoderm called somites

form on the outside of the neural tube,
become the vertebrae and associated
parts of the segmented body
 vertebrae

 muscles

 kidneys and reproductive structures

 heart and blood vessels

104
 Somites give rise to vertebrae
and muscles

examples of segmentation in humans

105
 Other organs are stimulated to form as a result, in part of
neural crest cell movements and differentiation
 Digestive tract formation starts as a simple tube separated
in the middle by the yolk stalk
 Separates into foregut and hindgut

 Liver, pancreas and trachea are hollow outgrowths from

the gut

106
 The trachea grows from the gut and lungs
develop from it
 The pharyngeal pouches grow laterally from
the pharynx
 Branchial grooves meet the pharyngeal
pouches and form branchial arches,
important in many structures of the head
 The pharyngeal and branchial grooves form
the gill slits and gills in aquatic vertebrates

107
 Mammalian Development
Human Prenatal Development

 Gestation lasts 266 days from fertilization to birth

 Development begins in the oviduct
 About 24 hours after fertilization, the zygote has

divided to form a 2-celled embryo

 The embryo passes down the oviduct by cilia and

peristalsis
 The zona pellucida (a vestige of the egg shell) has

dissolved by the 5th day, when the embryo enters

the uterus
 The embryo floats free for several days, nourished

by fluids from glands in the uterine wall

• At this point, it is called a blastocyst (same as
blastula) 108
 24 hrs
1 day

5 days

7 days

109
 The trophoblast is the outermost layer of cells
in the blastocyst
 The trophoblast forms the chorion and amnion
 The inner cell mass forms the embryo itself

110
 The Placenta
 The adaptation of placentation renders the large yolk
deposits in the egg nonessential. (there is still a yolk sac in
mammals but, it is empty)
 The placenta is the site of nutrient, gas, and waste
exchange
 It also secretes hormones that maintain pregnancy
 Trophoblast cells release human chorionic gonadotropin (hCG)
which signals the corpus luteum to enlarge and produce
progesterone FYI
 The placenta develops from the embryonic chorion and
maternal uterine tissue
 Chorionic villi are formed from the chorion, and project
into the endometrium of the uterus
 The umbilical cord, containing two umbilical arteries and
one umbilical vein connects the embryo and the
placenta
111
Development of the Placenta

112
12-day Human Embryo

113
 Organ Development

 Begins during the

first trimester
 Gastrulation occurs
during the 2nd and
3rd weeks, followed
by neurulation
(formation of the
neural tube)
 The heart beats
spontaneously after
3.5 weeks
 After the first two
months of Week 5
development, the
products of
conception are
114
called a fetus
 At the end of the first
trimester (first 3 months of
development)
 Fetus can be recognized as
a human
 ~56 mm long, and ~14 g
33-day embryo measuring 7
 The sexes can be x 3.2mm.

differentiated
 Ears, eyes becoming well-
developed,
 Skeleton starting to develop
 Notochord replaced with the
developing vertebral column
Human embryo at 40
 Moves, ‘breathes’, makes days.

sucking motions with thumb

115
 EvoDevo

 Ernst Haeckel noticed that

although adults of vertebrate
and arthropod classes look
very different, there is a
stage of development ,
occurring after gastrulation,
in which the embryos look
surprisingly similar.
 Ontogeny recapitulates
phylogeny
 What remains of this notion?
116
 The Phylotypic Stage

 Withina particular
phylum, embryos Amphibian

may differ in kinds

Bird
of eggs, cleavage
and organ Mammal

formation yet
share a common
post-gastrula
phylotypic stage. 117
 After vertebrates complete The Vertebrate
gastrulation and Phylotypic
neurulation that the 4
defining chordate features Stage
are visible
 Notochord
 Dorsal hollow nerve chord
 Gill slits
 Post natal tail
 The phylotypic stage of
embryo development
provides a flexible and
robust platform for
generating species
diversity.
118
 The phylotypic stage in vertebrates
represents an archetypal vertebrate body
plan.
 The cellular tactics can play upon the
archetypal body plan much like a
musician can play many different songs
on the same instrument.
 The answers lie in the transcriptional
signaling systems that are activated (or
deactivated) during development

119
 Mollusks

Annelids

Arthropoda

Echinoderms

Uchordates

Mammals

Aves

Reptiles

Amphibians
120
 What are the stages of development?

1. Pre-embryonic development - 1st week of

development after fertilization

2. Embryonic development – 2nd week after fertilization

until the end of the 2nd month

3. Fetal development – the 3rd through the 9th months of

development

4. Development after birth – stages of life including

infancy, childhood, adolescence and adulthood

121
 Pre-embryonic development
 Cleavage – cell division that increases the
number of cells

 Morula – compact ball of embryonic cells

 Early blastocyst – inner cell mass that

becomes the embryo covered by a layer of
cells that becomes the chorion

 Implantation – embryo embeds into the

uterus around day 6
122
123