Power Point 2.

3: Developmental Mechanisms
in Model Organisms

Migrating cell
Learning Goals
1. Describe mechanisms (cell specification,
axis formation, compartmentalization, etc.)
that are the foundation for the organized
development in model organisms.

2. Provide examples of developmental

specification processes and the genes
involved.
 What are some mechanisms
underlying development?
1. Cytoplasmic determinants: maternal substances (such
as mRNAs encoding transcription factors) in the egg
that influence the course of early development

2. Signaling molecules from neighboring cells

Sources of developmental information for the
early embryo

In (b) Certain signal molecules cause specific changes in gene expression

sending the cell down a specific developmental path; cells become different with
regard to structure and function.
Cell fate
Cells make developmental decisions by responding to
signals in their internal local environment (eg, cytoplasmic
determinants, signaling molecules).

Cells make developmental decisions within a temporal

and spatial framework
 C. elegans

Cell lineage of every somatic cell (959 cells in adult hermaphrodite) has been mapped
 C. elegans: P granules
Complexes of RNA and proteins
Initially distributed throughout the
embryo
Are repositioned to the posterior end
of the zygote and are found in the P4
blastomere, which gives rise to the
gametes

 https://www.youtube.com/watch?v=aVUWLSlMkxQ
 https://www.youtube.com/watch?v=M2ApXHhYbaw
Body Plan
Before the tissues and organs develop, the relative positions of
an animal’s head/tail, back/front and right/left are determined.

In frogs: A/P axis is determined during oogenesis

Dark melanin granules found in animal hemisphere
Yolk in vegetal side

In Frogs, D/V axis determined at fertilization

Cortical rotation: plasma membrane rotates toward the point of
sperm entry
 FROG

Cortical rotation enables molecules in parts of the vegetal cortex to

interact with molecules in animal cortex leading to specific gene expression
Chicks
A/P axis may begin to be established by gravity as the
eggs moves down oviduct

pH differences between the two sides of the blastoderm

cells lead to D/V patterns
 Pattern Formation
(insects)

Pattern formation: the development of a spatial

organization in which the tissues and organs of an
organism are all in the right place.

Begins in the early embryo when the major body axes are
formed: A-P, D-V, M-L

Often involves morphogenic gradients

 Morphogen
A morphogen is a substance governing the pattern of tissue
development, and the positions of the various specialized cell
types within a tissue.
It spreads from a localized source and forms a concentration
gradient across a developing tissue

Morphogen gradients generate different cell types in distinct

spatial order

Can be the products of genes ( eg., bicoid) or signaling

molecules (eg., Retinoic acid)
Pattern Formation
There are numerous genes involved in setting up axes (or
pattern formation) in animals!

The development of Drosophila is particularly well

studied
One Example: bicoid “two-tailed”
bicoid:
Homeobox gene; encodes for a transcription factor

Maternal effect gene: when it is mutant in the mother, the

offspring will have a mutant phenotype regardless of their
own genotype
bicoid

An embryo whose mother has two mutant copies

of bicoid has posterior structures at both ends

Thus, the bicoid gene product is essential for

setting up the anterior end of the fly
bicoid
bicoid mRNA is concentrated at the anterior end of the mature
egg

Once the egg is fertilized, it is transcribed into protein

The protein diffuses toward the posterior, resulting in a gradient.

bicoid binds to enhancers of other genes involved in pattern

formation, turning on genes that will direct cells to form
appropriate anterior structures.
 Gradient distribution of bicoid in a normal drosophila embryo
(more concentrated at the anterior end).
As the fly embryo develops…
Maternal mRNAs are destroyed so that the embryonic
program of gene expression can take over and
orchestrate further differentiation and development by
“refining” the original specifications.

 Embryonic genes encode for proteins that serve as

transcription factors (engrailed) and signaling molecules
(hedgehog, wingless); are involved in gene activation or
repression.
Major Classes of Developmental Genes
(drosophila)

Maternal effect genes - set up the A/P and D/V axis polarity

Gap genes - affect the development of a contiguous block of segments

Pair-rule genes - control the proper development of adjacent segments

(pairs)

Segment polarity genes - affect individual segments' polarity

Hox genes - affect the identity, characteristics of a particular segment

 Hox genes

Many drosophila pattern formation genes have homologues in mammals

Hox genes:
Expressed along the A/P
axis in the embryo in the
same order as their genes
are aligned on the
chromosome (3’5’)

Gene duplication and

divergence resulted
in vertebrates having 4
copies of the Hox gene
complex

Hox genes specify positional

identity, not a specific structure
Hox- homeobox
The products of Hox genes are known as Hox proteins.

Hox proteins are transcription factors as they are capable of binding

to specific nucleotide sequences on the DNA called enhancers
where they either activate or repress genes.

The same Hox protein can act as a repressor at one gene and an
activator at another.

The homeodomain is a 60 amino acid long DNA-binding domain

(encoded by the homeobox on the DNA).
Pattern formation in vertebrate
development (eg., limb)

Human embryo 6 weeks 9 weeks

Model System: Chick limb
Each component of a limb (a specific bone, or muscle)
develops at a precise location due to positional information

Limb bud: mesodermal tissue covered by ectoderm

Specific spatial patterns of Hox gene expression are

established early

Differential Hox gene expression cause specific cells in the

limb bud to react differently to positional cues
 Hox gene expression in limb

Different Hox genes are

“turned on” as limb
development progresses
(temporal, spatial refinement)
AER and ZPA
Two main organizing regions in limb
bud:

AER: apical ectodermal ridge

ZPA: zone of polarizing activity

AER
Consist of a thickened region of ectoderm at the tip of
the bud

 Involved in proximal-distal organization

Cells in AER secrete proteins in the FGF family (fibroblast

growth factor) that promote and maintain limb bud
outgrowth
 ZPA
 ZPA: mesodermal tissue located where the posterior side of
the bud is attached to the body.

Organizes the A/P axis; indicates “posterior”

Cells nearest to the ZPA give rise to posterior structures

(outer digit); farthest to anterior structures (thumb)

 ZPA cells secrete sonic hedgehog (SHH)

Figure 47.24a

Anterior
Limb bud

AER

ZPA
Limb buds
Posterior
50 m

Apical
ectodermal
ridge (AER)

(a) Organizer regions

Figure 47.25
EXPERIMENT
Anterior
New
ZPA
Donor Host
limb limb
bud bud
ZPA
Posterior

RESULTS

4 3

2
2

4 3
SHH
Is a signaling molecule and transcription regulator
involved in limb development (also, midline brain
structures, eye, teeth, and others).

Specifies digits

shh-/shh- yields a loss of digits

Also, if you add cells that express large amounts

of Shh, to the anterior region of the limb, result
will be “posteriorization” like transplant study in
previous slide
Drosophila Hedgehog Pathway

Shh
(vertebrates)

Gli1, 2, 3 (vertebrates)
 Summary of a few key genes in limb development
Hox genes: homeobox, transcription factors; specify the
“limb field” or “pre-pattern”

FGF: fibroblast growth factor, from AER cells, for limb outgrowth

SHH: Sonic Hedgehog; secreted signaling molecule, from ZPA, involved in

A/P patterning, specifies digits

WNT7: signaling protein found in dorsal limb ectoderm, involved in D/V

patterning (also neural tube development, cancer-colon)

BMP: bone morphogenesis protein; involved in interdigit programmed cell

death (regulates AER- FGFs )
 What are some mechanisms
underlying development?
The Cytoskeleton
What is the “Cytoskeleton”:
Network of proteins (actin filaments,
microtubules, intermediate filaments) in the
cytoplasm that provide the cell with structure
and shape.

Movement of parts of a cell can bring about

changes in cell shape or enable a cell to
Reorganization of the Cytoskeleton in Neurulation

Microtubules extend and

elongate the neural plate cells

Actin filaments at one end of the cell

contract; cells form a curved shape

Changes in cell shape result in a

hinge region where the neural
tube pinches off
Cell migration
Cell’s “crawl” within an embryo by extending and
retracting cellular protrusions.
Cell Crawling
Examples:
Predatory amoebae
Neural crest cells (migration from neural tube)
Macrophages, neutrophils (phagocytose material at site of
infection)
Osteoclasts (tunnel into bone)
Fibroblasts (rebuild damaged structures at injury sites
Cancer cells: cells crawl into blood vessels or lymph
Actin polymerization/depolymerization moves leading edge forward.
Contraction at back propels cell body forward. Back: contacts are
Disassembled. At front: new contacts made.
Cell migration
Also involves cell adhesion molecules and the ECM (extra
cellular matrix)
Extracellular Matrix (ECM)
Contains molecules (such as glycoproteins, eg: collagen,
fibronectin) that are secreted by the cell.

Integrins: receptor proteins found on the surface of cells

Bind to fibronectin and other ECM glycoproteins
Bind to proteins attached to the microfilaments of the
cytoskeleton
Transmit signals between the ECM and cytoskeleton
Example of a mechanism of migration
in a developing embryo

Fibronectin and other ECM proteins “communicate” with

integrins which affect cytoskeleton

Some cells in a developing embryo migrate along specific

pathways by matching the orientation of their
microfilaments to fibers in the ECM
 Another mechanisms underlying
development…
Apoptosis: programmed cell death
Reabsorption of a tadpole’s tail
Interdigital regions
Recap- Topic 2: Development
The process of fertilization has specific components.

Many organisms have shared (yet uniquely adapted) features during

development:
Cleavage, blastula formation, gastrulation, organogenesis

Gene expression and signaling (in a temporal and spatial manner), drive
cell determination and differentiation, giving rise to the body plan.

Mechanisms that underlie development include cell migration and

apoptosis