Development of the

Nervous System
Felipe | Fontanilla | Galang | Magpoc | Mariano | Medina
OVERALL STRUCTURE
AND CELL TYPES
Central Nervous System
-consists of the

● brain
● spinal cord

-consists of the
● Cranial nerves
● Spinal nerves
● Autonomic nerves
● and their associated Ganglia

Source: https://socratic.org

Peripheral Nervous System

The nervous system is formed from the
the neural crest, from epidermal
placodes and from axons growing from
cell bodies in the CNS
Nerve cells
Have numerous dendrites receiving
input from
other neurons.

source:https://www.youtube.com/watch?v=bfokof2YMVU

*in the CNS

Grey matter - mainly cells(cell bodies)
-also referred as brain nuclei
White matter- nerve axons
(whiteness from from high lipid
content of myelin sheath
Connections between neurons and
are called synapses

This is accomplished by:

1.)Release of neurotransmitter from presynaptic terminal
2.)opening of the ligand-gated channel in the postsynaptic terminal
3.)initiation of action potential
Types of Neurons
classified by
● morphology of axon and dendritic
tree
● Neurotransmitter used
Types of Neurons
classified by
● morphology of axon and dendritic tree
● Neurotransmitter used and their target tissue
○ Target tissue
■ Another neuron
■ Motor neuron(ex. Muscle, gland)
● From CNS to outer parts
■ Sensory neuron(sense organs)
● From outer parts to CNS
Other cells in nervous system
● Glial cells
○ Surround neurons and hold them in place
○ Supply nutrients and oxygen to neurons
○ Remove carcasses of dead neurons
 Glial cells
1.)Astrocytes 2.)Oligodendrocytes
○ Most numerous cell type in ● Cells of CNS that form
CNS the myelin sheath which
○ Structural and metabolic which are around the axons
functions ● NG2 Cells-precursors
○ Maintenance of blood-brain
barrier 3.)Schwann cells
○ Neurotransmitter uptake ● Equivalent of oligodendrocytes
○ Two Types In the PNS
■ Type 1 (Fibrous)
● Numerous in
white matter
■ Type 2(Protoplasmic)
● Numerous in grey
matter
 Glial cells
4.)Radial glial cell
● Present only during development
● Not really glia cell but make up a type
Progenitor cell.
● Present in lumenal surface of neural tube
and guides the radial migration of
daughter neurons
5.)Microglia
● phagocytic cells derived
from a primitive population
of embryonic macrophages
6.)Ependymal
● Simple cuboidal epithelial
lining of the ventricles and
and spinal canal
Retrograde transport
● shuttles molecules/organelles away from axon termini toward the cell
body.
● useful for neuroanatomists to trace pathways of axons.
○ Done by injecting a marker (such as horseradish peroxidase/HRP)
■ Taken up by axons transported to cell body.
 THE BRAIN
Appears from neural tube during early development.

FOREBRAIN
● Most anterior part
○ Telencephalon
■ Forms paired cerebral hemispheres
■ In lower verts, receptive area for olfactory organs
● Olfactory sensory cells from nasal placodes
■ In higher verts, where more complex associative and
and control behavior.
● In mammals- multilayered cerebral cortex
○ Neocortex
 THE BRAIN
Appears from neural tube during early development.

FOREBRAIN
● Posterior forebrain
○ Diencephalon
■ Produces the optic vesicles.
● Invaginate to form optic cups
○ Epidermis of optic cup- lens of eye
○ Inner layer-sensory retina
○ Outer layer-pigmented retina
■ Retina differentiates and sends signals to brain via
the optic nerve(in the optic stalk), terminate in the
midbrain
■ Forms thalamus-center of integration
■ the pineal body(epiphysis)-diurnal rhythms and source of melatonin
■ Pituitary- endocrine organ
 THE BRAIN
Appears from neural tube during early development.

MIDBRAIN
● Mesencephalon
○ Dorsal region- optic lobes
■ optic teca in non mammals
● Homologous to superior colliculus in mammals
○ Areas of reception of visual information
○ Also forms auditory lobes
 THE BRAIN
Appears from neural tube during early development.

HINDBRAIN
● Rhombencephalon
○ Dorsoanteriorly contains the cerebellum
■ For control of movement
○ Posteriorly contains medulla oblangata
○ Early embryo
■ Segmented to form 7 rhombomeres
● 1st forms cerebellum
● 5th and 6th are the otic placodes
○ Invaginate to form inner ear
■ Cochlea- organ of hearing
■ Semicircular canals- perception of motion and
control of balance
THE BRAIN

Neural tube’s lumen -persists in the ventricles of the brain and central
canal of spinal canal
Basic Set of Cranial Nerves
 The Spinal Cord
Consists of an inner ependymal layer,
● inner layer- grey matter-with most of the cell bodies
● outer layer - white matter- with mostly myelinated fibers
Grey matter
● Ventrally- motor neurons
● Dorsally- interneurons

● Commissural neurons- connect one of side of spinal cord to another

● Sensory neurons- contained in the dorsal root ganglia
○ Formed from neural crest

*At the level of the fore and hindlimb buds form networks
Called brachial plexus and lumbosacral plexus
-these nerves supply the limbs
 The Spinal Cord
Autonomic nervous system
divided into
● sympathetic
● parasympathetic
ANTEROPOSTERIOR PATTERNING
OF THE NEURAL PLATE
Initial Anteroposterior Pattern
Chapter 7 (Xenopus) & Chapter 9 (Chick)
Initial Anteroposterior Pattern
2 Domains

1. Anterior Part
● Anterior endoderm
● Prechordal mesoderm
2. Posterior Part
● Notochord
● Somites
Initial Anteroposterior Pattern
1. Anterior Part -- Goosecoid
● Moves towards animal pole
● Characterized by expression of transcription factor genes
(Goosecoid)
● Induce anterior type genes from ectoderm
● Otx2 (forebrain and midbrain)
● ag1 (cement gland)
● Anterior inductive activity because of secretion:
Cerberus (Inhibit Wnt, BMP, and Nodal-related factors)
Dickkopf (Inhibit Wnt)
Initial Anteroposterior Pattern
2. Posterior Part -- Homeobox genes
● Elongates by convergent extension
● Characterized by expression of different transcription factor
genes (Homeobox genes)
● Induce BOTH anterior and posterior type genes from ectoderm
● Otx2 (forebrain and midbrain)
● Hox genes (posterior)
● Posterior inductive activity because of secretion:
FGFs and Wnts (upregulate homeodomain transcription
factors encoded by cdx genes)
Summarized Initial Anteroposterior Pattern

Anterior Posterior

Characterized by gene
Goosecoid Homeobox genes
(Transcription factor gene)

BOTH anterior and posterior

Anterior type genes
type genes
Induce
Otx2 Otx2
ag1 Hox genes

Inductive activity due to Cerberus FGFs

secretion of Dickkopf Wnts
Anteroposterior patterning of the
Neural Plate
Anteroposterior patterning of the Neural Plate
● Early inducing factors produce a Central Nervous System
○ Divided into anteroposterior domains
○ REFINED by number of local interactions

Neural Induction

1. Forebrain
2. Posterior structures
3. Hindbrain
Overview
1. Forebrain
- Chordin (released from organizer)
- Inhibition of BMPs (Bone morphogenetic proteins)
- Induces a neuroepithelium with anterior specifications
- == Absence expresses Otx
2. Posterior structures
- Starts with action of posteriorizing factors (FGFs and Wnts)
- Induce expression of cdx genes
- Induce Hox genes
3. Hindbrain
- Retinoic acid
- Important posteriorizing function
Forebrain
Cerebral cortex
- Formerly thought that patterning was dependent on innervation from thalamus

o Mouse knockouts

- Even if innervation is lost – cerebral cortex showed initial pattern of

transcription factor domains

o Concluded that thalamus innervation REFINES the cerebral cortex

(which are established autonomously)
FGFs (Fibroblasts growth Factors)
● Inducing signal required for forebrain regionalization
● From anterior neural ridge
● First discovered in zebrafish as a region at extreme anterior of neural plate
● FGF8/FGF17/FGF18 – exists in mouse
○ When removed
- caused expansion of diencephalon
○ When grafted posteriorly
- induce ectopic telencephalon
○ Overexpression (in anterior)
- Shift boundaries of forming structures to posterior
- Introduction of structure in posterior cause duplication
Cortical Hem Region
● Posterior edge of developing cerebral cortex
● Where Wnts and BMPs are expressed
o Mouse knockouts for Wnts/Lef1
- Showed defects in posterior telencephalon
- Lack hippocampus

Wnts

● needed/required in posterior telencephalon formation

Emx2, Pax6, Couptf1, Sp8
● Transcription factor genes that are expressed in forebrain
in response to the signals of:
■ FGF
■ Wnt
■ BMP signals
● Show predictable changes of expression in
loss-of-function and gain-of-function mutants affecting
inducing factors
Isthmic Organizer
Isthmus

○ Constriction between midbrain and hindbrain vesicles

At open neural plate stage

○ Transcription factor genes (Otx2 and Lim1)

■ Expressed in forebrain and midbrain
■ Cells start to produce inducing factors
● Fgf8
● Wnt1
Isthmic Organizer
● anteroposterior signaling center
● regional specification of the neural tissue on either side
● can maintain itself after initial establishment (Fgf8)
● expression of Pax family in different regions:
○ Pax6 – expressed in fore- and hindbrain
○ Pax2 and Pax5 – expressed in midbrain and
rhombomere 1 region
○ Mouse knockout
■ lack midbrain and cerebellum
Isthmic Organizer
- Transplantation of isthmic region of chick embryo

Anterior

- Induces midbrain
- establishes a gradient expression of Engrailed 1 and Engrailed 2

Posterior

- induces rhombomere 1
- then cerebellum

activities shared by pure Fgf8 protein

Fgf8
● induce expression of
○ Engrailed (En)
○ Wnts
○ its own gene
■ Isthmic organizer can maintain itself after initial
establishment
Mouse Knockouts
● Wnt1 & En1
○ lack isthmus region and large parts of midbrain and
cerebellum
● Fgf8
○ Lethal
○ large posterior body defect
■ Hypomorphic allele of Fgf8
■ Zebrafish mutant acerebellar
Segmentation of The Hindbrain
Hindbrain
- Segmented
- Seven bulges (Rhombomeres) visible after neural tube
closure

Rhombomere 1

- Becomes cerebellum
Each subsequent pair of rhombomere contains
repeated set of motor nuclei and contributes
fibers to one cranial nerve:

Trigeminal (V) nerve from Rhombomere 2&3

Facial (VII) nerve from Rhombomere 4&5

Glossopharyngeal (IX) nerve from

Rhombomere 6&7
Every other rhombomere contributes to Neural
crest cells to one of branchial arches

Rhombomere 2 to 1st arch

Rhombomere 4 to 2nd arch

Rhombomere 6 to 3rd arch

Clonal Analysis

● Segmental character of rhombomeres

● Single cell is labeled
○ Progeny will cross between rhombomeres at an early stage but not at a
late stage

Clonal restriction

● arises because of aggregation of cells in the individual

rhombomeres
● due to alternating rhombomeres of ephrins and
eph-type receptors
Ephrin B-type ligands

● expressed on rhombomeres 2, 4, and 6

Ephs

● expressed on rhombomeres 3 and 5

** repulsion interaction creates boundaries **

● Overexpression of dominant negative Eph

- boundaries are not formed
In open neural plate stage

- rhombomeres open to change

- can be exchanged in position without affecting final pattern

In closed neural plate stage

- once rhombomere boundaries begin to constrict

- character can no longer be re-specified by transplantation
Notch system
- determines the actual formation of the boundary cells

In zebrafishes,

● Overexpression of Notch system

○ driven into boundary regions
● Overexpression of dominant negative Su(H)
○ excluded from the boundaries
Hox Genes
- Not expressed in forebrain and midbrain
- Important for anteroposterior patterning more
POSTERIORLY
- Upregulated by transcription factors
- Transcription factors upregulated by early gradients of

FGF, Wnt, and Retinoic acid

Hoxa2
- Expressed with its anterior limit at the future
rhombomere 1/ 2 boundary and expression in the
emergent neural crest cell is seen in rhombomere 4 not
in 2.
○ Knockout (mouse)
■ Homeotic shift of arch 2 towards arch 1
morphology
○ Overexpression (chick and xenopus)
■ Shift from arch 1 to arch 2 morphology
Krox20
● Zinc-finger protein expressed in rhombomere 3 & 5
● Expresses Eph-type receptors
● Does not emit neural crest stream
○ Mouse knockout
o Rhombomere 3 & 5 not formed
o 2/4/6 fuse
Retinoic acid
● Manipulations showed strong effects on the hindbrain
● From dietary vitamin A
● Forms a concentration gradient falling from posterior to
anterior across the hindbrain region
● Retinaldehyde dehydrase (RALDH2)
○ makes retinoic acid
● Cytochrome P450 (Cyp26)
○ Destroys retinoic acid
Quail embryos
● Almost retinoid free (depriving vit A)
● Has small hindbrains (Lacking rhombomere 5-7)
● Will grow if given vit A before early somite stage
● Loss of ventral neuronal types in the anterior spinal cord
Mouse Embryo
● RALDH2 knocked out
○ Die at mid-gestation
○ Lack Rhombomere 5-7

Xenopus Embryo
● Overexpression of dominant negative retinoic acid
receptor
○ Die at mid-gestation
○ Lack Rhombomere 5-7
Treatment of excess retinoic acid at E7.5
- cause Hoxb1 to be expressed in rhombomere 2 as well as the normal
expression in 4
- this alters the character of neural crest cells that come out of rhombomere 2
- converts first arch into a copy of the second arch

Teratogenic Effect
Spinal Cord
Spinal Cord
● Anteroposterior pattern is not so evident as in the brain
● Hox gene expression at various levels
● Anteroposterior arrangement of certain groups of motor
neurons
○ Lateral Motor Neuron Columns
○ Column of autonomic motor neurons
○ Electroporation of Fgf8 into column of terni
Lateral Motor Neuron Columns
- Supply the limbs
- Forelimb level – expression of Hoxc6
Column of autonomic motor neurons
- Column of terni (chicks)
- Lies in between the limb levels -- Expression of Hoxcc9
Electroporation of Fgf8 into column of terni
- Causes anterior shift of Hoxc6/Hoxc9 boundary
- Shrinkage of lateral motor column
DORSOVENTRAL PATTERNING
OF THE NEURAL TUBE
Spinal cord
Floor plate
● Composed of
columnar cells in
close proximity to
the notochord
● On each side is the
region containing
the motor neurons
Motor neurons
● Express a particular
combination of
Lim-type transcription
factors
○ Associated with the
group of muscles
Dorsal half of
spinal cord
● Dorsal sensory
relay interneurons
● Commissural
neurons
Roof plate
● Located in the
dorsal midline
Inductive Signals
Sonic hedgehog (Shh)
● Induces the floor plate and the motor neuron-forming
territory (at lower concentration)
● Shown by recombination experiments
● Floor plate and motorneurons will form
● Evidence that Shh is threefold:
○ Expressed in the notochord
○ Induce floor plate and motor neurons when applied to neural tube
○ Floor plate and motor neurons are lost in the mouse knockout of
Shh
Sonic hedgehog from the notochord induces expression of its own gene in
the floor plate. BMP from the epidermis induces the roof plate
The signals from the two centers induce a nested expression of genes that
later brings about the pattern of neuronal differentiation
Sonic hedgehog (Shh)
● Made by floor plate
● Contributes to the inducing role of the notochord
● Upregulates the expression of one set of transcription factor
genes (Foxa2, Nkx2.2, and Nkx 6.1)
● Represses the expression (Pax6 and Pax7)
Diagram of the initial gene expression domains and the corresponding progenitor domains in the
spinal cord. FP, floor plate; MN, motorneurons; p-domains are progenitor domains; V0-V3 are
interneuron domains
Diagram of the initial gene expression domains and the corresponding progenitor domains in the
spinal cord. FP, floor plate; MN, motorneurons; p-domains are progenitor domains; V0-V3 are
interneuron domains
Diagram of the initial gene expression domains and the corresponding progenitor domains in the
spinal cord. FP, floor plate; MN, motorneurons; p-domains are progenitor domains; V0-V3 are
interneuron domains
Diagram of the initial gene expression domains and the corresponding progenitor domains in the
spinal cord. FP, floor plate; MN, motorneurons; p-domains are progenitor domains; V0-V3 are
interneuron domains
Diagram of the initial gene expression domains and the corresponding progenitor domains in the
spinal cord. FP, floor plate; MN, motorneurons; p-domains are progenitor domains; V0-V3 are
interneuron domains
Diagram of the initial gene expression domains and the corresponding progenitor domains in the
spinal cord. FP, floor plate; MN, motorneurons; p-domains are progenitor domains; V0-V3 are
interneuron domains
Holoprosencephaly
● Human congenital defect
● Involves midline defects ranging in severity from the
presence of a single midline incisor tooth, to very severe
cases in which there is a single midline eye with a
proboscis above it
● Arises from a partial loss of function of Shh
○ Mutation in the SHH gene itself
○ Defects in cholesterol metabolism that prevent the addition of
cholesterol to the active form of the Shh molecule
Holoprosencephaly
● Can also arise from mutations in other genes
● Attributable to loss of function of Shh also show defects in
limbs and vertebrae
Dorsal midline
● Arises from the lateral
margin of the neural
plate, where it contacts
the epidermis
Epidermis
● Produces BMP4 and BMP7
○ Can elevate expression of Pax6 and Pax7 when applied to the
neural plate
● Roof plate expresses Bmp4
and -7 and the gene factor for
another TGF-β superfamily
member called Dorsalin
○ Have the activity of inducing
dorsal cell types such as sensory
relay neurons
The overall dorsoventral pattern of the spinal cord arises from the combined effects of the
gradients of Shh from the floor plate and of BMPs and Dorsalin from the roof plate.
NEUROGENESIS AND
GLIOGENESIS
Principles of Neurogenesis
Ventral Nerve Cord
● Arises from a Neurogenic region

Neurogenic Region
● Originates lateral to the mesoderm
● Mesoderm Invaginates → Midventral
● Contains a segmented reiterated pattern of Proneural Clusters
Proneural Clusters
● Defined by expression of bHLH transcription factor
● Position depends on the operation of the
anteroposterior & dorsoventral patterning system
● Produces:
○ Neuroblast
○ Epidermal cells

bHLH Transcription Factor

● Produced by Achaete-scute
complex (AS-C)
Achaete-scute Complex (AS-C)
● Produces bLHL transcription factor
● Activates the production of Delta

Delta
● Stimulates Notch Lateral
Inhibition System on adjacent
cells

Notch Lateral Inhibition System

● Represses expression of AS-C
● Controls the selection of the future Neuroblast
 Once the 1. Detaches
2. Sink into the Interior

Neuroblast is 3. Remained temporarily

connected by an Apical Stalk in

Formed the Plane of the Epithelium

Par Complex
● Phosphorylates components of the
intracellular trafficking system
● Repels Numb, Prospero, & Brat

Apical Side Basal Side

● Inherits the ganglion
● Inherits the apical stalk
mother cell
Primary Neurogenesis
 In Lower Vertebrates

First Neurons
● Arise from Neuroepithelium of
the open Neural Plate

Neuroepithelium
● Still one cell thick
● Appears stratified
Neurogenin
● Expressed in areas of Neural plate
● Give rise to first Primary Neurons
● Upregulation of NeuroD

NeuroD
● Highly expressed in Primary Neurons

Lateral Inhibition Mechanism

● Spacing of the Primary Neurons
❖ All Cells are competent to become Neurons
➢ Notch Signaling inhibits this tendency

❖ Overexpression of Delta
➢ Reduces the formation of Primary Neurons

❖ Overexpression of a Dominant Negative Delta

➢ Generate high density of Primary Neurons
Late Neurogenesis
In Mouse
2nd Half of Gestation
● Where most of Primary Neurogenesis occurs
● Neuroepithelium is more than one cell thick

Epithelial Structure
Apical Side
● Faces the lumen of the Neural Tube

Basal Side
● Faces the exterior
In Neuroanatomy
Apical Side
● Ventricular

Outer Surface
● Pial

At Early Stages
❖ All cells are Mitotic
❖ There’s an Interkinetic Migration
❖ A portion of Neuroepithelial cells transformed into Radial Glia
Radial Glia
● Progenitor cells for both Neurons & Glia
● Retains the apical Par Complex, Prominin-1, & blbp

In Vertebrate Neurogenesis
❖ Par Complex attract Numb rather than repelling it
❖ Numb proteins are retained
❖ Postmitotic neuron is depleted in Numb

In Mouse
❖ Numb-like proteins seems to maintain Radial Glia in proliferative mode
❖ Radial Glia → Astrocytes
➢ Depends on Cardiotrophin
Gliogenesis
 Glial Cell Formation
● Occurs after Neurogenesis
○ Radial Glia → Oligodendrocytes
○ Radial Glia → Astrocytes
Glial Cell
● Migrate considerable distance before assuming their final position

In Spinal Cord
Astrocytes
● Originates from p1, p2, & p3 regions
● Reelin
● Slit
○ Help to control subsequent migration and positioning
Oligodendrocytes
● Originate from the dorsal and the pMN regions

pMN Region
● Specified by a combination of transcription factors (Ngn2, Olig1 & -2)

In Late Gestation
❖ Downregulation of Ngn2
❖ Nkx2.2 domain expands dorsally
❖ Production of Oligodendrocyte precursor cells
Oligodendrocyte Precursor Cells
● Normal fate: Oligodendrocytes
● In culture: 2 Astrocyte (CNTF exposure)

NG2 Glia (Polydendrocytes)

● Using CSPG4
○ In White Matter: Self-renew & Oligodendrocytes
○ In Grey Matter: type 2 Astrocytes
DEVELOPMENT OF
NEURONAL CONNECTIVITY
Growth Cone
•Located at the tip of a developing axon
•Filopodia and lamellipodia
-Actin microfilaments extend; myosin-type motor proteins contract
-RhoA collapses growth cone; Rac, Cdc42, and neurotrophins
promote extension
•Cell body and proximal axon do not move
- Synthesis of new materials
Guidance Molecules
-Guide growth cones; some facilitate guidance and some antagonize
•Contact attraction and contact repulsion factors
•Long-range attraction and long-range repulsion
-Neuron to neuron; diffusible / via a concentration gradient from a long-range chemoattractant
-Moving chemoattractant changes direction of growth of axons
Neuronal Pathways
•Spaces, regions rich in suitable substrates (such as laminin), or regions
free from repulsive factors
•Pioneer neurons (embryo) make initial connections
•Fascicles – pre-existing axonal tracts
•CNS: Commissural neurons
-Grow down the ventral midline (netrins and Shh from the floor plate)
-NgCAM causes crossing to dorsal side
Neurotrophins and Cell survival
•Neuronal death by apoptosis during postnatal growth of the CNS
•Survival of neurons depend on:
1.Growth factors which function as survival factors
- NGF group of neurotrophins, some FGF’s, hepatocyte growth
factor, GDNF, CNTF, etc.
2. Absorbing enough neurotrophin from target organ
Axonal Competition
•Reshuffling of connections (muscles)
•Several neurons to one myofiber one neuron to one myofiber, but
with greater number of synapses (neuromuscular junctions)
•Depends on electrical activity
-Neuromuscular junctions are strengthened by muscle excitation (active);
weakened if inactive
Neuronal Connections in the Visual System
•Visual field – area which objects can be seen as you focus your eyes on a central point
•Light from visual field Retina Retinal neurons Optic nerve Optic tecta (Brain)

•One-on-one (topographic) relationship

-Between retina and optic tectum
Chemoaffinity Hypothesis (Sperry)
•Specificity of connections
•Xenopus experiment
- Preferred hierarchy of connectivity; slight differences between models but
principles are the same
•Retinal fibers have to get to the tectum
- Gradients of cell-surface molecules between retina and tectum
-Ephrin A – anteroposterior; controlled by Engrailed
-Ephrin B - dorsoventral
Later Refinement
•Mammals:
- Birth: receptive fields for both eyes in the visual cortex are mixed
-Visual stimuli: ocular dominance columns (stripes)
oDo not form if neuronal activity is blocked
- Anterograde transport: Retinal cells Midbrain Cells Cortex (ocular dominance columns)

•Same principle with neuromuscular junctions:

- Simultaneous: adjacent points on retina of one eye; strengthened
- Not simultaneous: same visual point viewed through different eyes; weakened

From Simplicity to Complexity

Early stages do not need neuronal activity or visual experience; later refinement needs an input of
visual experience
The Neural Crest
What is the Neural Crest?
•It is a population of cells which is derived from the neural
fold
•They are present in the dorsal part of the neural tube;
•Soon migrate away from the neural tube
Neural crest forms a variety of cell types
•Neurons, ganglion of the sensory and autonomic systems
•Adernal medulla & Calcitonin cells of the thyroid
•Pigment cells
•Skeletal tissues of the head
•Part of the cardiac outflow tract
Establishing the fate of the neural crest
•Two ways:
•1) Grafting of segments of the neural tube
•2)In-vivo labelling
•DiL
•Fluorescent dextran
Domains of the Neural crest
The neural crest is divided into distinct anteroposterior domains:
1)Cephalic crest-Source of head mesenchyme
•Cartilage
•Bone
•Cranial Ganglia
2)Trunk crest-two pathways of migration
•Dorsolateral Pathway-passes through epidermis and somites
•Ventral pathway-passes through sclerotome
Domains of the Neural crest
3)Vagal Crest
●

4)Sacral crest
Formation of the neural crest
Clonal descendants can include:
•Epidermal
•Neural
•Neural crest cells
•Combination of the three
Formation of the neural crest
•Environment that promotes formation of the neural crest
•Removal of neural fold
•Neuroepithelium joined directly to epidermis
•BMP4-IMPORTANT PROTEIN IN NEURAL CREST CELL FORMATION

•Molecular pathway
Molecular pathway for determination
•Linear sequence
•BMP signaling
•Expression of Msx1 and 2 homeodomain transcriptional repressors
•Snail and slug(zinc-finger transcriptional repressors)
•Sox9 and 10
•Overexpression, inhibition and epistasis
Molecular pathway for determination
•Overexpression of msx factors will induce expression of snail
•Overexpression of snail will induce expression of sox10
•Overexpression of sox10 will induce expression of slug

•Injection of morpholinos or domain swap inhibit neural crest

formation
Molecular pathway for determination

Msx Snail Sox10 Slug

Formation of neural crest
•Glucocorticoid-inducible version of slug
•If induced early, Block neural crest formation
•If induced late, block cell migration
Formation of neural crest
•Sox10 is initially expressed in the whole crest but soon is replaced by
sox9 in the cranial crest
•Sox9- cartilage differentiation in the mesoderm
•Ablation of sox9 inhibits formation of cranial skeleton without
affecting the formation of the trunk formation
•Snail inhibits the production of e-cadherin and components of tight
junctions
Migration of cells in the Neural Crest
•Depends on a variety wide variety of extracellular matrix components and
proteases
•Matrix metalloproteinase-degradation of ecm proteins during organogenesis
•Adams-enables penetration of the cells
•Fibronectin
•Laminin
Migration of cells in the Neural Crest
•The migration of the trunk crest of the ventral pathway is inhibited by the
posterior sclerotome which leads to the preferred migration route the
anterior sclerotome

•Ephb3 is expressed by crest cells

•Ephb1 and 2 is expressed by the posterior sclerotome
•Leads to actin depolymerization
Commitment and Differentiation
•Experimented by interchanging segments of the neural tube and
examining the fate of the neural crest cells
•Only the cranial crest can generate skeletal tissues
•Trunk crest can form cartilage when grafted into the facial primordia
•Any region of the neural crest will form the parasympathethic &
sympathetic n.s. and sensory ganglia
Commitment and Differentiation
•Vagal crest-cholinergic(parasympathetic n.s.)
•Sacral crest-adrenergic(sympathetic n.s.)

•If interchanged, the cell types formed is appropriate for the new position of the
graft
Commitment and Differentiation
•Individual crests that leave the neural tube early are multipotent
•Sensory neurons, pigment etc.
•Those leaving the neural tube late are more likely to populate only a
single cell type
•melanocytes
Commitment and Differentiation
•The final differentiation of the cells, in some instances, depend on the
environment(inducers)
•Dorsal root ganglia-exposure to bdnf
•Schwann cells-neuregulin
•Smooth muscle-tgfb
•Autonomic neurons-BMP2 or 4