Module 4:

Cell Signalling in Development

Lecture 3: Ectoderm/Neural Developlment

MCB*3010
Prof. Gordana Scepanovic
Mar. 12, 2026
Lecture Outline
1) Find it, lose it, move it
2) The structures of the ectoderm
3) Modes of neurulation & how the neural tube is
established
4) Revisiting Wnt signalling
5) D/V patterning of the neural tube
6) A/P patterning of the neural tube
7) Neuronal positioning
Studying embryonic development
Find it: use microscopy to visualize tissues, cells, molecules that mediate a process and their
dynamics.

Lose it: remove the tissues/cells/molecules under study (using genetics, drugs, etc). Does
the process still happen normally? à necessity.

Move it: reproduce the tissues/cells/molecules under study in a different location (using
transplantation, genetics, etc.). Does the process happen in the new location? à sufficiency.
Patterning the vertebrate embryo:
the ectoderm

• The ectoderm has three major responsibilities:

1. Form the epidermis (the outer layer of the
skin).
2. Form the neural plate (which will become
the neural tube, a precursor to the central
nervous system - CNS).
3. Form the compartment residing between
epidermis and the CNS, the presumptive
neural crest (these generate pigment cells
and the peripheral nervous system - PNS).
Patterning the vertebrate embryo:
the ectoderm

What signals determine whether

the ectoderm will form
epidermal vs neural tissues?
 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).
Establishing neural tissue

• The “default fate” of ectoderm is to become

neural tissue.
• Three key molecules “protect” the neural
fate by blocking BMP signalling:
• Chordin
• Noggin
• Follistatin
Overview of the nervous system

• The nervous system is divided into the

central nervous system (CNS) and the
peripheral nervous system (PNS).
• The CNS comprises the brain and
spinal cord; and
• the PNS consists of all the neural
structures outside the CNS.
The development of the nervous system

ectoderm

neuroectoderm

neural tube

neurons
The development of the nervous system
neural tube
neural plate

mesoderm notochord

epidermis

Neurulation: the process where the three ectodermal regions

become physically and functionally distinct from one another.
An embryo undergoing these processes is called a neurula.
Modes of neurulation

Principal modes of neurulation:

1. Primary neurulation, where cells
surrounding the neural plate direct the
neural plate cells to proliferate,
invaginate into the body, and separate
from the surface ectoderm to form an
underlying tube.
2. Secondary neurulation, where the
neural tube arises from the aggregation
of mesenchyme cells into a solid cord
that subsequently forms cavities that
coalesce to create a hollow tube.
The development of the neural tube

• Bending of epithelial cells can form several

important developmental structures,
including the neural tube.
• The neural plate must invaginate to form the
neural tube.

How are these cells connected

to move in unison?
 The development of the neural tube

• The development of the neural tube

includes precisely orchestrated
morphogenetic movements, including:
1. The thickening of the
neuroectoderm.
2. Apical constriction of neural plate
cells via spatially regulated
actomyosin contraction.
3. Secondary bending of the tube to
support closure at the anterior region
promotes a more circular structure.
4. Closure of the tube is established by
Rho remodeling cell-cell adhesions.

Kinoshita et al. (2008) MBoC

 The development of the neural tube
Vehicle-treated control Y-27362 treated
• Blocking RhoA activity by targeting its
downstream effector, RhoA, with Y-
27362 inhibits MLCK phosphorylation,
thus reducing local myosin II
contraction.
• Treatment with Y-27362 after
neuroectodermal thickening disrupted
the formation of the neural tube.

What would happen if you

RhoA ROCK local myosin II injected Y-27362 after the first
MLCK
contraction bending of the neural plate?
Y-27362
Kinoshita et al. (2008) MBoC
Wnt signalling
• Wnt signals through two different pathways:
• Canonical Wnt
• Wnt binding to receptors on the cell
surface (Frizzled) promotes the
stabilization of the protein β-catenin.
• When β-catenin accumulates, it
translocates into the nucleus and
activates transcription of many
genes.
• Involved in the generation of neural
progenitors and specialized
subtypes.
• Non-canonical Wnt
 Wnt signalling

• Wnt signals through two different pathways:

• Canonical Wnt
• Non-canonical Wnt
• Wnt binding to receptors on the
cell surface (Frizzled) promotes the
recruitment of the phosphoprotein
Disheveled, which can activate
RhoA signalling to influence
cytoskeletal remodeling.

Akoumianakis et al. (2022) Nature Review Cardio.

Determining the signals required for
neural tube development

RNA interference (RNAi): uses

small RNA molecules to promote
the sequence-specific degradation
of target RNAs and prevent them
from being translated into proteins.
The two main RNAi-based
strategies are small interfering
RNA (siRNA) for temporary
effects, and short hairpin RNA
(shRNA) for lasting results.
 Determining the signals required for
neural tube development
Frizzled shRNA
treated cells

invagination
neural tube
control Frizzled shRNA

• Wnt signalling can be specifically blocked on one side of the embryo by injecting half of
the cells of an 8-cell stage embryo with shRNA against Frizzled.
• When Wnt signalling is knocked down, Rho does not accumulate, and thus there are no
cytoskeletal rearrangements to promote neural tube invagination.

Kinoshita et al. (2008) MBoC

The development of the neural tube

• Failure in neural tube closure can result in

severe birth defects. For example:
• Anencephaly is the absence of a
major portion of the brain, skull, and
scalp,
• Encephaloceles are abnormal
openings of the skull that permit the
herniation of intracranial contents, and
• Spina bifida where the spine doesn't
fully develop.
The development of the nervous system

Major neural
signalling center
The role of the notochord in
neural tube patterning
• Find it: Shh is found in the notochord
and the developing floor plate,
corresponding to the areas being
patterned.
• Lose it: If the notochord is removed, the
adjacent neural tube fails to develop
floor plate cells. This shows that the
notochord is necessary.
• Move it: Transplanting a notochord—or
even just cells secreting Shh—to a new
location causes the induction of an
ectopic floor plate and motor neurons.
This proves Shh is sufficient to dictate
this development.
D/V patterning of the neural tube
dorsal
• Morphogens are secreted from the ventral or
dorsal side relative to the midline of neural tube.
ventral • Shh – ventral
• Floor plate (notochord)
• Mesencephalon
• Motor Neurons
• Dopaminergic Neurons
• BMP and WNT – dorsal
• Roof plate
• Sensory Neurons – Neural tube (CNS)
• Neural Crest – PNS
• Paraxial Mesoderm – muscle
D/V patterning of the neural tube

• The concentration gradient established

between Shh, Wnt and BMP result in
expression of different transcription
factors in each part of the neural tube
• This gives rise to different functional
groups of neurons:
• Sensory neurons (relay sensory
information, heat, pain…) will form on
the dorsal side, and
• Motor Neurons (involved in muscle
contraction) form on the ventral side.
D/V patterning of the neural tube

• BMP and Shh contribute to neuronal

diversity by each activating specific
Paired Box (Pax) transcription
factors.
• Shh activates ventral Pax
transcription factors (Pax6) in the
ventral neural tube
• BMPs activate the dorsal Pax genes
(Pax 3 and Pax 7) and represses
ventral PAX genes (Pax 6) in the
dorsal spinal cord.
D/V patterning of the neural tube

• Neural tube patterning depends

on a combination of amount and
duration of morphogen
exposure.
• For example, increasing doses
of Shh results in a loss of
ventral patterning genes, e.g.
Pax7. Similarly, increasing the
duration of Shh exposure will
also repress Pax7.
A/P patterning of the neural tube

• Anterior-posterior neural tube

patterning results in two key structures:
• Brain: prosencephalon (forebrain),
mesencephalon (midbrain),
rhombencephalon (hindbrain)
• Spinal Cord
A/P patterning of the neural tube

anterior • The bi-directional gradient of Wnt

and FGF allows the brain to
dorsal ventral
develop in a distinct manner from
posterior the spinal cord.
Wnt
• Like segmentation of the
Drosophila embryo, the hindbrain
Shh is also segmented in structures
FGF
called rhombomeres. Unique
neurons and nerves arise from
each rhombomere.
• Hox genes establish the different
rhombomeres.
 A/P patterning of the hindbrain
anterior posterior
• Hox gene expression in the hindbrain is
regulated by retinoic acid (RA)
control receptors.
• A gradient of RA exists from the
posterior to the anterior axis.
RA gradient • High levels of RA activate posterior Hox
genes and repress anterior Hox genes.
• Thus, RA is considered a teratogen, an
RA treatment agent responsible for congenital
disruptions.
Development of the brain

• Neurons are born in the central cavity of

the neural tube and then migrate outwards
in order to populate the brain.
• While the original tube is only a single
layer, the rapidly dividing neural stem cells
quickly establish three layers:
• The highly proliferative ventricular
zone,
• The mantle or intermediate zone, and
• The marginal zone (the outer cells of
the tube).
Development of the brain

• Newly born neurons must migrate

from the ventricular zone to the
anterior, marginal zone, which will
ultimately form a brain structure.
• Radial glial cells are specialized
neuroectoderm cells that function as
stem cells that can gives rise to new
neurons and glia. They can also form
the scaffold on which neurons can
migrate.
 Development of the brain

• Radial glial processes guide migrating

neurons and support their migration
by establishing and maintaining
interactions. These include:
• Integrin-mediated cell-cell
interactions;
• N-cadherin cell-cell adhesions;
• Gap junction adhesions; and
• Branched migration, utilizing
dynamic filopodia structures.

Meyerink et al. (2020) Cells

Development of the brain

• Once the neural tube closes, the

apical surface of the neuroepithelium
will border the internal cavity.
• Radial glia maintain a polarized
morphology spanning the apicobasal
axis of the central nervous system
(CNS). This supports the orientation
and positioning of new cells.
Development of the brain

• New neurons are continuously

expanding outward.
• Thus, the first neurons born travel less
distance to the site of terminal
differentiation. The next round of
neurons can then migrate past the
first to sequentially grow the brain.
• This results in the formation of layers
in the newly developing neocortex.
Development of the brain

How do neuronal axons

avoid its own dendrites?

Alison Barth
Development of the brain

• Dscam encodes a membrane-

adhesion protein that prevents
dendrites from the same neuron from
touching one another.
• When two dendrites from the same
Dscam-expressing neuron come in
contact, they repel each other.
• Thousands of Dscam isoforms are
generated through RNA splicing to
ensure that each neuron acquires a
unique identity.

Alison Barth
 Studying neurogenesis
• The development of optogenetics and
chemogenetics has revolutionized systems
neuroscience by allowing for targeted excitation
and inhibition of specific neuronal subpopulations
in discrete brain regions.

Optogenetics uses a light-sensitive ion channel that

is expressed in targeted cells, allowing for neuronal
depolarization or hyperpolarization with pulses of light.

Chemogenetics involves the expression of

DREADDs (designer receptors) that are targeted to
specific neurons, which can be activated by “designer”
ligands locally or systemically to induce neuronal
activation or inhibition.

Maya Peters Kostman

Studying neurogenesis: optogenetics

• A gene encoding channelrhodopsin can be introduced into a subpopulation of

neurons in a mouse. Channelrhodopsin can be optogenetically controlled, so that in
response to blue light, channels open, the neuron is depolarized and activated.
• When the light is switched on, the mouse immediately becomes aggressive and
attacks; when the light is switched off, its behavior immediately returned to normal.
In summary
You should be able to:

1) Describe how to incorporate the “find it, lose it, move it” method to study
developmental cell biology.
2) Compare the structures formed by the ectoderm.
3) Understand how the neural tube is generated during neurulation.
4) Explain how D/V and A/P patterns are established in the neural tube.
5) Describe how neuronal migration contributes to brain growth.
6) Understand how optogenetics may be used to study neuron function.
 Module 4:
Cell Signalling in Development

Next time:

Lecture 4 - Review