Chapter 8 Principles of Development

Donna May dela Cruz-Papa College of Science University of Santo Tomas

Primary Organizer
Hans Spemanns Research only tissue to induce growth Research in 19201930s centered on: how one tissue influenced the fate of another: induction 1916 - Spemann noted the capacity of tissue from the dorsal lip of the gastrula-transformed tissue 1921-22 - Repeated delicate experiments showed it induced a 2 embryo All of the knowledge for constructing a complex egg must be in the nucleus and cytoplasm

Advances in Molecular Biology

2nd golden age of embryology Spemann organizer cells secrete proteins: 1. Noggin 2. Chordin 3. Follistatin Secretion of certain molecules trigger or repress the activity of a combination of genes in nearby cells Similar proteins occur in both invertebrates and vertebrates These scientific breakthroughs ushered in the new science of Evolutionary Developmental Biology

Early Concepts: Preformation Vs. Epigenesis

Preformation - miniature adult being present in the sperm or egg, waiting to unfold (others claim they see the adult) - young animal is merely unfolding the structures that are already there Kaspar Friederich Wolff (1759) - NO preformed chick in the early egg - Undifferentiated granular material became arranged into layers - Layers thickened, thinned, and folded to produce the embryo Epigenesis is the concept that the embryo contains building materials that are assembled

Development
series of progressive changes 1. when a fertilized egg divides mitotically 2. Specialization - occurs as a hierarchy of devtal decisions 3. Cell types do not unfold but arise from conditions created in preceding stages 4. Interactions become increasingly restrictive; each stage limits developmental fate 5. With each new stage, cells lose the option to become something differentit becomes determined 6. Both cytoplasmic localization and induction cause this feature

2. Fertilization
A. Initial Event Fertilization - union of male and female gametes - provides for recombination of paternal and maternal genes, restoring the diploid number - activates the egg to begin development B. Oocyte Maturation - contrast with sperm Sperm - eliminates nearly all cytoplasm and condenses its nucleus Egg - grows in size by accumulating yolk - contains much mRNA, ribosomes, tRNA & elements for protein synthesis Morphogenetic determinants direct the activation and repression of specific genes in the egg The egg nucleus grows in size, bloated with RNA and is called the germinal vesicle. Preparations in the egg occur during the prolonged prophase After meiosis resumes, the egg is ready to fuse its nucleus with the sperm nucleus

Fertilization & Activation

marine invertebrates: especially sea urchins Contact and Recognition Between Egg and Sperm Sea urchin sperm first penetrate the jelly layer before contacting the vitelline envelope Egg-recognition proteins on the acrosomal process bind to species-specific sperm receptors on the vitelline envelope

Prevention of Polyspermy
Fertilization cone - forms where the sperm contacts the vitelline membrane Important changes in the egg surface block entrance to any additional sperm Polyspermy - entry of more than one sperm - would cause a triploid nucleus In the sea urchin, an electrical potential rapidly spreads across the membrane; this is the fast block - followed by the cortical reaction

Cortical Reaction
enzyme-rich cortical granules below the egg membrane fuse with the membrane granules release contents in space between the membrane and vitelline envelope Then, the reaction Water rushes into space elevates the envelope Lifts away all sperm bound to it. Except for one One cortical granule enzyme causes the vitelline envelope to harden, becoming the fertilization membrane

Fusion of Pronuclei and Egg Activation

After fusion - sperm disconnects from its flagellum Enlarged sperm nucleus (male pronucleus) migrates inward to contact female pronucleus
diploid nucleus Nuclear fission

Sea urchins (12 mins); mammals (12 h) The Egg is now a zygote DNA and protein synthesis undergoes a burst of activity, using a supply of mRNA in the egg cytoplasm

Fertilization initiates reorganization of cytoplasm and repositions determinants that begin development and cleavage

Cleavage & Early Development

A. Blastomeres Embryo undergoes cleavage/segmentation
- convert large cytoplasmic mass into small maneuverable cells

No cell growth occurs

only subdivision until cells reach regular somatic cell size

Polaritya polar axisestablishes the direction of cleavage and differentiation

Patterns of Cleavage
Cleavage - orderly sequence of cell divisions The pattern of cleavage is affected by:
a. quantity and distribution of yolk present, and b. genes controlling the symmetry of cleavage

Vegetal pole - yolk-rich end of the embryo while the other end is the animal pole During each division, a cleavage furrow is visible in the cell

1st to 3rd cleavage

How Amount and Distribution of Yolk Affects Cleavage Isolecithal yolk - eggs with very little yolk and the yolk is distributed evenly
a. In such eggs, cleavage is holoblastic b. The cleavage furrow extends completely through the egg

Isolecithal eggs - widespread and seen in: echinoderms, tunicates, cephalochordates, molluscs and mammals Note: Cleavage is slowed in the yolk-rich vegetal pole

 Mesolecithal eggs - moderate amount of yolk concentrated in the vegetal pole

Animal pole contains cytoplasm and very little yolk Holoblastic cleavage but cleavage is retarded in yolk-rich vegetal pole Cleavage furrow slow- vegetal pole thus cleavage is faster in the animal region

Telolecithal eggs - much yolk concentrated at the vegetal pole

Actively dividing cytoplasm is confined to a narrow discshaped mass on the yolk Cleavage is partial or meroblastic; - furrow does not cut through the heavy yolk Birds, reptiles, most fishes and a few amphibians have telolecithal eggs

 Centrolecithal eggs - large mass of centrally located yolk

Cytoplasmic cleavage is limited to a surface layer of yolk-free cytoplasm; yolk-rich inner cytoplasm is uncleaved They have meroblastic cleavage (Fig 8.8) Insects and many other arthropods have centrolecithal eggs

Amount of yolk affects developmental mode

a. In most animals, a mother does not directly nourish embryonic development but has provisioned the egg with yolk b. The amount of yolk is related not only to cleavage pattern, but also to whether a larval stage occurs during development c. Animals in which the zygote is telolecithal generally have direct development - occur when there is enough yolk to support growth as juveniles (in reptiles and birds) d. Species with isolecithal or mesolecithal zygotes generally have indirect development - animals where the larval stage is between embryo and adult

What Can We Learn from Development?

Biologists study development for different reasons a. to understand how a single cell (the zygote) can produce the variety of body parts in an organism b. search for commonalities among organisms The division of the 32 multicellular animal phyla into groups is simplified by an understanding of their development

Overview of Devt Following Cleavage Blastulation

Cleavage creates a cluster of cells called the blastula In most animals, these cells are arranged around a fluid-filled cavity called the blastocoel

 Blastula stage - consists of a few hundred to several thousand cells Blastula formation, with a single germ layer, occurs in all multicellular animals In all animals other than sponges, development continues to form one or two more germ layers Germ layers formed during development ultimately produce all structures of the adult body

Gastrulation and Formation of Two Germ Layers

Gastrulation - results in the formation of a second germ layer - typically involves invagination (pushing inward) of one side of the blastula Invagination forms a new internal cavity the archenteron or gastrocoel Blastopore - opening into the cavity The gastrula has an outer layer of ectoderm and an inner layer of endoderm. The only opening into this embryonic gut is at the blastopore.

Some animals retain a blind, or incomplete gut, but most have a complete gut with a second opening, the anus.

Formation of a Complete Gut

The inward movement of the archenteron continues until the end of the archenteron reaches the ectodermal wall of the gastrula. An endodermal tube, the gut, is surrounded by the blastocoel The endodermal tube has two openings, the blastopore and a second opening formed by the merging of the archenteron tube with the ectoderm.

Formation of Mesoderm, a Third Germ Layer

Animals with two germ layers - diploblastic Most animals add a third germ layer - triploblastic Mesoderm 3rd germ layer between the endoderm & ectoderm - can form in two different ways: 1. proliferation of cells from near the lip of the blastopore into the space between the archenteron and the outer body wall 2. by the pushing of the central region of the archenteron wall into the space between the archenteron and the outer body wall Regardless of method of mesoderm formation, initial mesoderm cells arise from endoderm

Formation of the Coelom

body cavity surrounded by mesoderm Coelomic cavity appears within the mesoderm by either schizocoely or enterocoely (same coelom)
The method by which the coelom forms is an inherited character and is important in grouping organisms based on developmental characters

Upon completion of coelom formation, the body has three tissue layers and two cavities

Suites of Developmental Characters

Two major groups of triploblastic animals 1. protostomes (insects, sea urchins, shellfish,
earthworms)

2. Deuterostomes (vertebrates, starfish) **identified by a suite of four developmental characters

Deuterostome Development
Cleavage Patterns Radial cleavage - embryonic cells are arranged in radial symmetry around the animal-vegetal axis Regulative development - fate of cell depends on interactions with neighboring cells
Early in the development of these embryos, each cell is capable of producing an entire embryo if separated from the other cells

Fate of Blastopore
Deuterostome embryos develop a complete gut blastopore anus 2nd opening mouth Coelom Formation
Enterocoely - mesoderm & coelom form at the same time - gastrulation begins with one side of the blastula bending inward to form the archenteron - pouch-like compartment pinches off to form a mesodermally bound space surrounding the gut

 Radial cleavage is characteristic of the Deuterostomia, echinoderms, hemichordates & chordates bilateral cleavage - egg is defined by unequal cytoplasmic components(Fig 8.12) - Ascidians (tunicates) demonstrate this cleavage pattern. discoidal cleavage (telolecithal eggs) - there is a large mass of yolk in each egg; cleavage is confined to a small disc of cytoplasm - Early cleavage furrows carve the disc into a single layer of cells (blastoderm)

Examples of Deuterostome Development Variations in Deuterostome Cleavage

 Rotational cleavage 1st cleavage plane - aligned with the animal vegetal axis 2nd cleavage 1 blastomere divides meridionally (through the animal-vegetal axis)while the other divides equatorially (perpendicular), rotated 90 degrees to the first Early divisions may be asynchronous and possess odd numbers of cells below the 2-4-8-16 series that would occur with synchronous division 3rd - cells form a tightly packed cluster stabilized by outer cells with tight junctions, the trophoblast trophoblast - form the embryonic portion of the placenta type of cleavage is present in mammals and is slower than in other animal groups

Examples of Deuterostome Development Variations in Deuterostome Gastrulation

sea stars - gastrulation begins with the flattening of the vegetal area of the blastula to form the vegetal plate - archenterons elongates toward the animal pole and its anterior end expands to form two pouchlike coelomic vesicles
- Ectoderm - epithelium of body surface & nervous system - Endoderm - epithelial lining of the digestive tube - Mesoderm - muscular & reproductive system, peritoneum & calcareous plates of the sea stars endoskeleton

 Frogs - radial cleavage, movements during gastrulation - influenced by the large mass of yolk Gastrulation in amphibians begins with invagination

Birds & reptiles Primitive streak forms the anterioposterior axis and center of early growth blastoderm has 2 layers: epiblast and hypoblast with a blastocoel between them Epiblast sheet moves toward primitive streak and over the edge, migrating One stream of cells moves deeper, displacing midline hypoblast, and forms endoderm Surface cells form ectoderm

mammalian gastrulation Epiblast move medially through the primitive streak into the blastocoel cells migrate laterally through the blastocoel to form mesoderm and endoderm mammals utilize nutrients from a placenta

Reptiles, birds & mammals share a common ancestor whose eggs were telolecithal; all inherited this gastrulation pattern and mammals then evolved isolecithal eggs with a telolecithal pattern

Protostome Development
Cleavage Patterns Spiral cleavage - protostomes Blastomere cleave obliquely to the animal-vegetal axis. Upper layer of cells appears offset relative to lower layer Mosaic devt is characteristic of most protostomes. (Fig 8.10) - cell fate is determined by the distribution of morphogenetic determinants, in the egg cytoplasm

 Protostomes are so named because the blastopore becomes the mouth. Coelom Formation a band of mesoderm forms around the gut before a coelom forms If a coelom exists, it is formed by schizocoely (splitting) Endodermal cells move by ingression (entering) into the space between the archenteron walls and the ectoderm These cells divide and produce mesodermal precursors; the proliferating cells become the mesoderm. Some protostomes, like flatworms, are acoelomate In others, mesoderm lines only one side of the blastocoel, leaving a fluid-filled cavity around the gut called a pseudocoelom.

Fate of Blastopore

Examples of Protostome Development

Two clades of protostomes 1. Lophotrochozoan protostomes
- contains segmented worms, molluscs - Some members of this clade employ a whorl of tentacles known as a lophophore and a trochophore larva

2. Edcysozoan protostomes
- includes arthropods, roundworms.. - name of the clade refers to ecdysis (taking it off or stripping off)

Examples of Protostome Development Variations in Protostome Cleavage

Superficial cleavage - centrally located mass of yolk restricts cleavage to the cytoplasmic rim of the egg

Mechanisms of Development
Nuclear Equivalence 1. Roux-Weismann Hypothesis a. An earlybut wrongexplanation of differentiation was based on division of nuclear material along the cell lineage b. Early embryologists saw this as an explanation of differentiation; genome gradually became broken down into smaller units c. Hans Driesch separated the two-celled sea urchin stage; both developed into normal larvae but this did not disprove eventual progressive modification

Hans Spemann Disproves Roux-Weismann Hypothesis, Discovers Gray Crescent

Spemann used human hair to almost tie-off a salamander zygote The nucleus was on one side; only cytoplasm on the other The nucleated side divided many times: only when one nucleus wandered across did the cytoplasmic side divide Its normal development showed no nuclear material had been lost after many nuclear divisions Occasionally the nucleated side only developed into a ball of belly tissue; it was missing the pigment-free gray crescent area that appears at fertilization. This disproved the Roux-Weismann Hypothesis and showed that cytoplasm in the gray crescent contained essential information

Spemanns experiment demonstrated that every blastomere contains sufficient genetic information for the development of a complete animal

Methods of Differential Cell Differentiation

Except for insects, cells become committed to particular fates from cytoplasmic segregation of determinative molecules during cleavage, and from interaction among neighboring cells (induction)

Cytoplasmic Specification
Cytoplasmic components are unevenly distributed in a zygote Different components contain morphogenetic determinants that control commitment to cell type Different determinants are partitioned among different blastomeres by cleavage and determine cell fate.

 Some tunicate species have different colored types of cytoplasm a. Yellow cytoplasm gives rise to muscle cells. b. Gray equatorial cytoplasm produces the notochord and neural tube. c. Clear cytoplasm produces larval epidermis. d. Gray vegetal cytoplasm gives rise to the gut.

Embryonic Induction
Induction - capacity of some cells to evoke a specific devtal response in other cells a graft of the dorsal blastopore lip could induce ectoderm to form a neural tube Dorsal lip was the primary organizer because it was the only tissue to induce growth Spemann called this primary induction the first inductive event

Cells that have differentiated act as inductors for adjacent undifferentiated cells Timing is critical; primary induction sets in motion secondary induction. The sequence includes cell movement, changes in adhesion, and cell proliferation

Gene Expression During Development After fertilization, proteins are translated from stored mRNA transcribed from maternal genome In many animals, maternal mRNA directs protein synthesis through cleavage and to mid-blastula stage After this, protein synthesis switches from maternal to zygotic control as the nucleus transcribes its own mRNA

Homeotic Genes and Hox Genes

Gene expression is regulated to ensure orderly devt Mutations of homeotic genes in fruit flies revealed they controlled overall body plan of legs, wings homeobox - 180-nucleotide DNA sequence occurs in most animals - master genes that control expression of subordinate genes Homeotic genes are remarkably similar across diverse species; Proteins coded by homeobox genes contain a highly conserved 60-amino acid sequence: the homeodomain. - they switch subordinate genes on or off

 Mice and humans - 4 clusters of homeobox-containing genes on separate chromosomes - all are homologues of the fruit flys homeotic genes Hox & homeobox genes - play a role in shaping individual organs and limbs (Fig 8.18, 8.19) New limb buds can be induced to grow from the side of a chick by implanting a bead soaked in fibroblast growth factor (FGF) helps establish a proximodistal axis one of the three axes that guide limb devt (Figure 8.20)

Evolutionary Devtal Biology

Zoologists have long looked to embryology for clues to evolutionary history. Evolutionary developmental biology has contributed several concepts to our understanding, but many questions remain. 1. Are the body plans of all bilaterally symmetric animals fundamentally similar? a. Insects and amphibians share a similar control of dorsoventral patterning, except that one is upside down relative to the other b. The French naturalist Geoffrey Hilaire (1822) - vertebrates were essentially inverted invertebrates c. His idea was rejected, but zoologists are reconsidering whether protostomes and deuterostomes are simply inverted with respect to each other 2. Can the anatomy of extinct ancestral species be inferred from the developmental genes shared by their descendants? a. Similar dorsoventral patterning in protostomes and deuterostomes suggests a common ancestor with similar patterning b. The same can be said for the similarity of HOM/Hox clusters in insects and chordates 3. Instead of evolution proceeding by the gradual accumulation of numerous small mutations, could it proceed by relatively few mutations in a few developmental genes? a. The induction of legs or eyes by a mutation in one gene suggests that organs can develop as modules. b. This raises the question of whether entire limbs or organs could be lost or gained during evolution as the result of one or a few mutations. c. The existence of developmental genes that could bring about major changes would challenge Darwins theory of gradualism.

Vertebrate Development
Common Vertebrate Heritage
One outcome of shared ancestry in vertebrates is the similarity of postgastrula embryos - all vertebrate embryos share: dorsal neural tube, notochord, pharyngeal gill pouches with aortic arches, ventral heart and postanal tail. - similarity is extraordinary considering the variety of eggs and developmental patterns

Amniotes and the Amniotic Egg

Amniotes are a monophyletic grouping of vertebrates - embryos develop within the amnion (membranous sac that provides an aquatic environment and protects it from shock) Amniotic egg has 4 extraembryonic membranes including amnion Yolk sac 1st membrane and pre-dates the amniotes by millions of years; it is extraembryonic and is discarded Allantois sac from the hindgut & holds metabolic wastes during devt Chorion - lies beneath the eggshell; encloses the rest of the embryo With growth ; allantois + chorion fuse to form the chorioallantoic membrane, a provisional lung

Mammalian Placenta and Early Mammalian Devt

Mammals inherited the amniotic egg but retained it in the mothers body Monotremes are examples of primitive mammals that lay eggs. Marsupials, embryos develop but do not take root in the uterus; they climb out and enter an external abdominal pouch Placental mammals represent 94% of the Class Mammalia - maternal oviduct had to evolve to develop a uterus to house the embryo

Monotremes
Absence of teats (modified sweat glands sucked by youngs) Lay eggs With cloaca Testes within abdomen (absence of scrotal sac) No pinna of ear Platypus (ductbill), Echidna (spiny anteater)

Young nursed in marsupium (abdominal skin pouch) Yolk sac serve as placenta Teats opening into marsupium Geographically isolated in Australia Ex. Kangaroo, koala, wallaby, opossum, Tasmanian wolf

Marsupialia

Placental mammals

Development of Systems and Organs

Germ Layers Germ layers should not be confused with germ cells (eggs and sperm) Germ layers do not alone determine differentiation but rather the position of embryonic cells

Derivatives of Ectoderm: Nervous System & Nerve Growth

Ectoderm thickens to form a neural plate Edges of neural plate fold up to create an elongated, hollow neural tube Neural crest cells pinch off from the neural tube. It is unique to vertebrates and was important in evolution of the vertebrate head and jaws Ross Harrison (1907) - cultured nerve cells; each axon grows from one cell

Derivatives of Endoderm: Digestive Tube and Survival of Gill Arches

During gastrulation, the archenteron forms as the primitive gut. This endodermal cavity eventually produces the digestive tract, lining of pharynx and lungs, most of the liver and pancreas, thyroid and parathyroid glands and thymus. During development, endodermally-lined pharyngeal pouches interact with overlying ectoderm to form gill arches In fish, gill arches become gills. Gill arches remain as necessary primordia for a variety of other structures in terrestrial vertebrates. 1st arch and its endoderm-lined pouch form the upper and lower jaws. 2nd, 3rd & 4th pouches become the tonsils, parathyroids and thymus

Derivatives of Mesoderm: Support, Movement and the Beating Heart

With an increase in size and complexity, mesodermally derived structures take up a greater proportion. Muscles arise from mesoderm along each side of the neural tube. The mesoderm divides into a linear series of somites (38 in humans). The splitting, fusion and migration of somites produce the: 1) axial skeleton, 2) dermis of dorsal skin, and 3) muscles of the back, body wall and limbs. Limbs begin as buds from the side of the body; projections become fingers and toes

Mesoderm gives rise to the embryonic heart

Guided by underlying endoderm, two clusters of precardiac mesodermal cells move to either side of the gut. These clusters differentiate into a pair of double-walled tubes that fuse into a single thin tube. Primitive heart begins beating on the second day of the 21-day incubation period; there are no blood or vessels at this time. Twitching becomes rhythmical as the ventricle and atrium develops; each chamber has a faster intrinsic beat. A specialized sinoatrial node in the sinus venosus eventually takes charge as pacemaker. The circulatory system becomes critical to deliver food from the yolk and return wastes to extraembryonic tissues.