Module 4:

Cell Signalling in Development

Lecture 1: Patterning-Tissue Development

MCB*3010
Prof. Gordana Scepanovic
Mar. 5, 2026
Lecture Outline
1) Processes of development
2) Cell specification
3) How patterning is established in the embryo
4) Patterning the Drosophila embryo
5) Patterning the vertebrate embryo
Every multicellular organism started its life
as a single cell.
A zygote begins with multiple rounds of cell divisions
that can then be specialized for different functions.

[Link]/watch?v=7Q9VyHJ1l2Q
Essential processes of development
On a fundamental level, Haeckel is correct…

Embryogenesis is conserved!

• The basic “machinery” of

development is the same in
all animals.

• Thus, homologous proteins

often perform the same role
in different species.

Romanes' 1892 copy of Ernst Haeckel’s famous drawing.

Basic animal body plan
Lateral
Medial
• Many animals have clearly defined axes:
Lateral
Dorsal • an anteroposterior axis, with mouth and
brain anterior and anus posterior;
Anterior
Ventral
• a dorsoventral axis, with back dorsal and
belly ventral; and

• a mediolateral axis (also referred to as

animal-vegetal), with medial (vegetal)
internal and lateral (animal) external.
Posterior
Conserved mechanisms establish the basic
animal body plan

Step 1: A fertilized egg (or zygote) rapidly divides or cleaves.

• At this stage the cell does not grow and its genome is inactive.
• Maternal Protein and RNA is degraded.
Conserved mechanisms establish the basic
animal body plan

Step 2: Once the embryonic genome becomes activated, cells cohere to

form the blastula
• The blastula – can be solid or hollow (if fluid filled, will form the
rudimentary gut).
Conserved mechanisms establish the basic
animal body plan

Step 3: A complex rearrangement of cells, called gastrulation, will form the

three germ layers:
• Ectoderm (will give rise to the epidermis and the nervous system);
• Endoderm (will form the gut tube and its appendeges i.e. lung, liver etc.); and
• Mesoderm (will give rise to the muscles, connective tissues, blood etc.).
Conserved mechanisms establish the basic
animal body plan

Blastocysts contain pluripotent cells that have

the potential to give rise to almost any of the cell
types of the adult body.

What cell type cannot be made

by pluripotent stem cells?
The developmental potential of cells becomes
progressively more restricted.
Blastomeres are cells of the inner cell mass of the
blastocyst. This includes human embryonic stem
cells (hESCs).

• Pluripotent cells can form mesoderm, endoderm,

ectoderm, and germ cells.

• Totipotent cells can form mesoderm, endoderm,

ectoderm, germ cells AND placenta.

• As cells develop, they lose pluripotency and take

on more restricted fates. Cell determination
starts early, progressively narrowing cell fate.
The developmental potential of cells becomes
progressively more restricted.

Why don’t cells go

backwards to take on
more pluripotent states?
Simple signals can generate complex
cell patterns

• Cells expressing an extracellular signalling cue will direct a new cell fate in their
neighbours through inductive signalling.
• Inductive signals include cell memory and combinational signalling.
Morphogens as inductive signals

A morphogen is a diffusible chemical

substance that acts over long distances to
instruct cells to adopt suitable fates for tissue
patterning by forming an activity gradient.

• The four major families of morphogens include:

1. The Fibroblast Growth Factor (FGF) family
2. The Hedgehog family
3. The Wnt family
4. The TGF-β superfamily
How do the simple morphogens form complex
animal body patterning?

• The initial patterns are

established in small fields of cells
and refined by sequential
induction as the embryo grows.
• Different animals use different
mechanisms to establish their
primary axes of polarization
(including anteroposterior,
dorsoventral and left–right axis.
How do the simple morphogens form complex
animal body patterning?

But what if all the cells in

a tissue are the same?
Can a pattern arise
within a tissue where the
cells are all the same?
How do the simple morphogens form complex
animal body patterning?

Latereral inhibition is a type of cell–cell

interaction whereby a cell that adopts a
particular fate inhibits its immediate neighbours
from doing likewise.

• Lateral inhibition can generate patterns of

different cell types.

• It is often mediated by the Notch

signalling pathway.
The Notch Signalling Pathway
• The Notch signaling pathway is a
highly conserved, direct cell-to-cell
communication mechanism that involves
a receptor (Notch) on one cells that
binds membrane-bound ligand (Delta)
on an adjacent cell.
• When one cell generates a slightly
higher Delta signal, it will activate Notch
on the neighbouring cell, and inhibit cell
specialization.
• Therefore:
• Active Notch = stay progenitor
• Inactive Notch = differentiate
Body patterning is established throughout
embryogenesis

• Most animals pass through

similar stages of development:
fertilization, cleavage,
gastrulation, organogenesis,
birth/hatching, metamorphosis
and gametogenesis.
• Embryogenesis includes all
of the stages of development
between fertilization and birth
(or hatching).
Fertilization: symmetry breaking

• Polarization is the first step of spatial patterning.

• Some animals (i.e. Drosophila or the frog Xenopus) have eggs with clear asymmetries
that establish the mediolateral (animal-vegetal) axes.
• In vertebrates, the site of fertilization establishes the dorsoventral axis.
• Fertilization leads to microtubule reorganization.
Lessons from Drosophila on patterning

The Nobel Prize in

Physiology or Medicine for
1995 was awarded to
Edward B. Lewis, Christiane
Nüsslein-Volhard and Eric
Wieschaus for their
discoveries concerning “the
genetic control of early
embryonic development”.
Lessons from Drosophila on patterning

• Cell populations are specified for

developmental fates in the early embryo
by location.

• Developmental fate is dependent on the

asymmetric distribution of
transcriptional regulators.
Lessons from Drosophila on patterning

Waves of cell division in the syncytium • The Drosophila egg has an established
anterior-posterior prior to fertilization.
• Insects go through a syncytial stage where
signals can diffuse readily through the
shared cytosol.
Lessons from Drosophila on patterning:
polarized expression of mRNA

In Situ Hybridization (figure A) is a technique

that allows for the precise localization of a
specific segment of nucleic acid or mRNA
within a histologic section.

How does the mRNA

profile compare to the
protein? Why?
 Lessons from Drosophila on patterning:
regulators of A-P patterning
• The fate of different subpopulations in an
embryo are determined by the presence or
absence of different transcriptional regulators.
• Segmentation genes will divide the embryo
into A-P segments. These include:
• Egg polarity genes: 1st expressed, divide
time
the AP region
• Gap genes: 2nd expressed, coarse AP
subdivision.
• Pair-rule genes: 3rd expressed, subdivides
segments made by gap genes on AP axis.
• Segment polarity genes: 4th expressed,
gives polarity to each segment.
Gheisari et al. (2020) Mech. of Dev.
 Lessons from Drosophila on patterning:
regulators of A-P patterning

How do we know what

time these genes do?

Gheisari et al. (2020) Mech. of Dev.

Lessons from Drosophila on patterning:
regulators of A-P patterning
Mutations in segmentation genes result in:

• Egg-polarity genes: a lack of

anterior structures.
• Gap genes: elimination of one or
more segments of the embryo.
• Pair-rule genes: series of
deletions affecting alternate
segments, resulting in half the total
number of segments.
• Segment polarity genes: number
of segments does not change, but
each segment lacks polarity.
 Lessons from Drosophila on patterning:
regulators of A-P patterning
GAP PAIR-RULE SEGMENT- Mutations in segmentation genes result in:
GENE GENE POLARITY GENE
• Egg-polarity genes: a lack of
anterior structures.
• Gap genes: elimination of one or
more segments of the embryo.
• Pair-rule genes: series of
deletions affecting alternate
segments, resulting in half the total
number of segments.
• Segment polarity genes: number
of segments does not change, but
each segment lacks polarity.

Niisslein-Volhard & Wieschaus (1980) Nature

Lessons from Drosophila on patterning:
regulators of A-P patterning
• Key transcriptional regulators are
expressed in a sequence.

• The collective presence of these

regulators promote spatially restricted
expression of additional transcriptional
regulators (sequential induction!).

• These actions control expression of

very important genes in development
(HOX genes) that define specific cell
types.
Lessons from Drosophila on patterning:
regulators of A-P patterning – Wnt & Hh
• Post cellularization, segment polarity genes that encode the signal
transduction proteins required for WNT and Hedgehog (Hh) signaling
are activated.
• Wnt and Hh are synthesized in different bands of cells to serve as
signalling centers within each segment.
• Hh regulates Wnt, Wnt indirectly regulates Hh through Engrailed.

Wnt
Lessons from Drosophila on patterning:
regulators of A-P patterning – Hox genes
Segment-polarity gene Engrailed
expression pattern.
• Initial patterning is unstable, and
yet the fundamental organization
persists in complex body
structures.
• The transient pattern established
by egg-polarity, gap, and pair-rule
genes is remembered by segment-
polarity and Homeotic selector
(Hox) genes.
• Hox genes are controlled by
Wnt and Hh.
Lessons from Drosophila on patterning:
regulators of A-P patterning – Hox genes
• Homeotic mutations (Hox
mutations) transform parts of the
body into structures appropriate for
other places.
• Transformation can only occur
between structures of similar
general types (i.e. changing one
limb to a different limb or changing
one segment into another).
• Local expression of Hox genes
gives regional position information,
that represents an intrinsic signal
of the cells’ location.
Lessons from Drosophila on patterning:
regulators of A-P patterning – Hox genes

What would a
mutation of ALL the
Hox genes look like?
Lessons from Drosophila on patterning:
regulators of A-P patterning – Hox genes
• Hox genes are expressed according
to their order in the hox complex.

• Their order along the chromosome,

mirrors their order of activity along
the AP axis, suggesting sequential
gene activation.

• This could result from changes in

chromatin structure and the
sequential opening and closing of
heterochromatin as it moves along a
chromosome.
Lessons from Drosophila on patterning: regulators of
A-P patterning – Trithorax & Polycomb

• Trithorax and Polycomb proteins enable the Hox complexes to

maintain a permanent record of positional information.
• Both are chromatin remodeling complexes which function via DNA
methylation and histone acetylation (epigenetic regulation).
• Polycomb keeps chromatin closed in regions where HOX genes
are not yet expressed.
• Trithorax, keeps chromatin open once a Hox gene is on by
blocking Polycomb action.
Lessons from Drosophila on patterning:
regulators of D-V patterning

• In the early embryo, there is a

gradient of nuclear-localized Dorsal.
• The amount of Dorsal protein
present in the nucleus spatially
determines whether additional genes
are expressed.
• Most dorsally, Dpp (TGF-β family
member) will be switched on,
followed by Sog and Twist. Together
these proteins will interact to create
the D-V territories.
Patterning the vertebrate embryo
• The origins of the embryonic axes and
the three germ layers in the frog can be
traced back to the blastula (fate
mapping):
• The endoderm derives from the
most vegetal blastomeres
• The ectoderm from the most
animal, and
• The mesoderm from a middle set.
• Within each of these territories, the
cells have diverse fates according to
their positions along the D-V axis of the
later embryo.
Patterning the vertebrate embryo:
regulators of A-V patterning
• TGF-β family patterns the AV axis.
• A high concentration near the vegetal-pole stimulates
endoderm formation.
• Two proteins Nodal and Lefty (TGF-β family
members) are secreted from the vegetal pole.
• Nodal is a TGF-β activator that acts locally
(because it diffuses slowly),
frog embryo
• Lefty antagonizes Nodal, and acts distal to the
site of secretion (diffuses rapidly)
• Thus, the concentration gradient of TGF-β signalling
establishes a pattern of progenitors – endoderm,
mesoderm, ectoderm along AV axis.
Patterning the vertebrate embryo:
regulators of D-V patterning

• A gradient of inhibition of TGF-β family members

patterns the DV axis.
• Bone morphogenetic proteins (BMPs – a
subfamily of the TGF-β super-family) are
secreted throughout the embryo.
• The dorsal signalling system releases two
proteins, Noggin and Chordin, that block
frog embryo
BMP signalling when their own concentrations
are high enough. This gradient therefore
blocks BMP on the dorsal side.
 Patterning the vertebrate embryo

• Putting it all together:

neural • High BMP + low Nodal results in
dermis
epidermal tissue (on the ventral side).
ectoderm
• Low BMP + low Nodal results in neural
tissue (dorsal side)
• This is an example of sub-patterning,
where from the ectoderm you create more
specialized tissues (epidermal and neural).
Patterning the vertebrate embryo: blastoids

Models of human embryos (blastoids) at the blastocyst stage.

Mallapaty (2024) Nature News
In summary
You should be able to:

1) Explain the essential processes involved in development.

2) Understand the initial steps of establishing the basic animal body plan.
3) Discuss pluripotency.
4) Understand how patterning arises within tissues.
5) Describe the morphogens involved in patterning the Drosophila embryo.
6) Explain how these patterns are maintained over time.
7) Compare and understand patterning in the vertebrate embryo.
 Module 4:
Cell Signalling in Development

Next time:

Lecture 2 - Endoderm/Mesoderm-Development