A: NEURAL CREST

CELLS
AND B: AXONAL
SPECIFICITY

Gilbert, Chap 13
A: The neural crest
 Transitionally structure
 Cells from neural crest migrate to become numerous
different cell types
 4 functional domains
Regions of the neural crest
1. The cranial neural crest
 Migrates into branchial arches
 Forms bone and cartilage of the face and neck
 It also produces pigment and cranial nerves

2. The cardiac neural crest cells= Somites 1-3

 Critical in making the division between the aorta and
the pulmonary artery
3. The vagal neural crest = Somites 1-7
 Forms the parasympathetic nerves of the gut

4. Neural crest cells of the trunk = somite 6 to tail

 Make the sympathetic neurons and a subset of these
(somite 18-24) form the medulla portion of the adrenal gland
1.2 Mechanisms of trunk neural crest
migration
1.2.1 Emigration from the neural tube
Three important interactions,
1) The RhoB protein establishes the cytoskeleton conditions
that promote migration
2) Slug protein activates factors that dissociate the tight
junctions between cells
3)The loss of N-cadherin that links these cells together

All migrating cells express HNK1.

Determinants of migration
 Extra cellular matrix and soluble factors
1) Promoters of migration = fibronectin, laminin,
tenascin, various collagen molecules and
proteoglycans
2) Impede migration = ephrin proteins
3) Chemotactic and maintenance factors
Stem cell factors allow the continued proliferation of
neural crest cells that enter the skin and may serve
as an anti-apoptosis and a chemotactic factors
 Trunk neural crest cell differentiation
Pluripotency of Trunk Neural Crest Cells

 Pluripotency = the ability to differentiate into any of

several different cell types depending on location
 However some neural crest cells are committed
very soon after leaving the neural crest
Final differentiation of trunk neural crest cells
 Determined by environment
 Become either A) adrenomedullary cell or a B)
sympathetic neuron

A) Adenomedullary cells (Figure next slide)

Glucocorticoids act at two places in the pathway;
1) Inhibit factors that promote neuronal differentiation
2) Induce enzymes found in adrenomedullary cells

B) Sympathetic neurons (Figure next slide)

Sequential exposure to basic fibroblast growth factor (FGF2)
and nerve growth factor (NGF)
 NGF competent
cell
NGF
Sympathetic
neuron
FGF 2 Inability to respond to
Bipotent glucocorticoids
precursor cell

Glucocorticoids
Pluripotent
neural crest cell
Inhibition of neural Glucocorticoids
differentiation
Promotion of chromaffin
specific enzymes

Chromaffin
(adenomedullary) cell
3. The cardiac neural crest
 Cardiac neural crest cells are pre-committed to
the development of cardiac cells
 Congenital defects of the heart are
often associated with defects of the parathyroid,
thyroid and thymus usually linked to defects in the
migration of cells from the neural crest
B: Neuronal specification and axonal specificity

 Axons of neurons also migrate

 Neurons produces axons that extend for meters
 Axon has a locomotory apparatus (in the growth cone)
 Responds to similar types of signals as migrating cells
 Signals are more specific than those for cell migration
1. The generation of neuronal diversity

Vertebrates form a dorsal neural tube

 by blocking a BMP signal
 Specification through the Notch-Delta pathway
Specification of the type of neuron

Controlled by the position of the neuronal precursor

within neural tube
 Eg: Ventrolateral margin = the motor neuron
Two periods of sonic hedgehog (Ssh) signalling
1. Early period - ventrolateral region are instructed to
become ventral neurons (Ssh from notochord)
2. Later period - ventral neurons are instruct to
become a motor neuron (Ssh from floor plate cells)

Ssh specifies motor neurons by inducing certain

transcription factors at different concentrations
Target specificity- Neuron innervates a specific target eg: thigh,
forelimb
Anterior –posterior specification of the neural tube is regulated
primarily by the hox genes (see later limb development)
Within a specific region, age regulates motor neurons specificity
 Younger cells migrate to the periphery
 Pass through neurons that have differentiated earlier in
development
 Early born neurons secrete factors such as retinoic acid that
cause the younger neurons to express new transcription factors
 Due to different birthdays and migrating patterns motor neurons
form the 3 major groups

 Cell bodies of motor neurons projecting into a single muscle are

called a pool
 3 larger columns according to their targets
Column of Terni (CT) -sympathetic ganglia
Lateral motor column (LMC) - limb musculature
Medial motor column (MMC) - axial muscles
2. Pattern generation in nervous system

Functioning of brain depends

 (i) on differentiation
 (ii) positioning of neurons
 (iii) but also specific connections the cells make amongst
themselves and their peripheral targets
AXONS - 3 main steps
1) Pathway selection – Axons travel along a path leading a particular
region of the embryo
2) Target selection - In specific region, recognises and bind to a set of
cells and form stable connections
3) Address selection Refining of initial patterns each axon binds to a
small subset of its possible targets

1 and 2 are independent of neuronal activity

3rd involves interaction between several active neurons and converts
the overlapping projections into a fine tuned pattern of connections
 2.1 Cellular adhesion & contact guidance
 Determined by environment that growth cone encounters
 Either specific signals or by
 Haptotaxis = the phenomena of migrating on an adhesive
gradient
 Growth cones migrate on surfaces that are more adhesive than
their surroundings eg.adhesive glycoprotein - laminin
 2.3 Contact guidance by specific growth cone
repulsion

 In addition to specific adhesion there is also specific repulsion

 Ephrin guides the growth cone by selective repulsion They are

especially important when the axon needs to make turns, does not
grow in a straight line
2.4 Guidance by diffusible molecules

Netrins and their receptors

 Netrin-1 is synthesised and secreted by floor plate

cells
 Netrin-2 is synthesized in the lower region of the
spinal cord but not the floor plate
2.5 Target selection
 Once the neuron has reached its target tissue it becomes
responsive to various proteins that are produced by the target
cells
 A group of these proteins, the neurtropins include nerve growth
factor (NGF), brain-derived neurotrophic factor (BDNF)
neurotropin 3 (NT-3) and NT4/5
 Each neurotopin can promote the growth of some axons to its
source while inhibiting other axons
2.6 Address selection: Activity-dependent
development
 Axon contacts its target it forms a synapse
 The construction of a synapse involves several steps
 Surface eg: laminin, fibronectin
 Selective repulsion: e.g Ephrin
 Soluble factors: eg: Netrin-1 and Netrin-2
 Responsive factors secreted by target cells:
E.g. nerve growth factor (NGF),
brain-derived neurotrophic factor (BDNF)
neurotropin 3 (NT-3) and NT4/5
 Example: Muscle (detailed)
Differentiation of motor neuron synapse
with muscle. Parts (E) and (G) lower
magnification to give an overview of the region
where the axon meets the muscle.
(A) Growth cone approaching a developing
muscle cell (B) the axon stops and forms an
unspecialized contact on the muscle surface.
Agrin released by the muscle causes
clustering of the acetylcholine receptors near
the axon
(C) Neurotransmitter vesicles enter the axon
terminal and an extracellular matrix connects
the axon terminal to the muscle cell as the
synapse widens. This matrix contains a nerve
specific laminin.
(D) Other axons converge of the synaptic site

(E) Overview of muscle innervation by several axons

(seen in mammals at birth)

(F) All axons but one is eliminated. The remaining axon

can branch to form a complex junction with the
muscle. Each axon terminal is sheathed by a Schwann
cell process and folds form in the muscle cell
membrane

(G) Overview of the muscle innnervation several

weeks after birth. Muscle derived laminin specifically
binds the growth cones of the axons and may act as a
stop signal for axonal growth
2.7 Differential survival after innervation:
Neurotrophic factors
 Neurotrophins regulate neuron death (apoptosis)
 Secreted by target tissue
 Removal of target tissue neuron death
 NGF (Nerve Growth Factor) = sympathetic+sensory neurons
 GDNF (Neurotrophin glial cell line derived neurotropic
factor) = midbrain dopaminegic neurons (Parkinson’s disease)

 Clinical trails of neurotrophic factors for the possible alleviation

of spinal cord injuries (NGF), Parkinson’s disease (GDNF), and
Alzheimer’s disease (NGF, GDNF)
2.9 Fetal neurons in adult hosts

 In 1976 fetal rat brain tissue transplanted into brain

of a newborn rat
 Neurons connected with correct regions of host brain
 Recently specific niches of cells have been identified
Parkinsons disease
 In Parkinsons, dopamine producing neurons of substantia nigra
are destroyed
 Axon terminal degenerates
 In 1990 Lindvall transplanted human neural cells from
foetuses (8-9 weeks) into brain of a Parkinson’s patient
 5 months later, levels of dopamine were normal
 No rejection due to the blood brain barrier
 ETHICAL ISSUES
 MPTP/Parkinson's disease and animal models

 70’s, student, B Kidston (USA) became curious about drugs

 Being a chemistry major, he to synthesized his own drugs
 Interested in a synthetic analog of Demerol (MPPP)
 Set up a laboratory in basement of his parents home
 He synthesized & used MPPP for several months without incident
 On a day the product caused a pronounced unexpected burning
sensation, and three days later, he began to experience
bradykinesia severe enough that he was admitted to hospital
 Parkinsons disease (PD) was diagnosed
 Treatment with L-Dopa improved his symptoms
 NIH was notified
 Drug analysis - MPPP and MPTP, an unexpected by-product
 Toxicity of MPTP was confirmed in animals
 Later in 1982 six young people in California suddenly manifested a
PD-like syndrome
 Doctors were baffled as patients were too young, and the symptoms
occurred too abruptly to be true PD
 Later it was found that these patients were all drug addicts
 Had taken a synthetic heroin (White China)
 Manufactured by a local drug dealer in his garage
 Found to be contaminated with MPTP
 From both incidents MPTP was identified as the responsible agent
 Led to the development of an animal model to study PD
 (ScienceDirect 1700 articles)
 Fig. 4. Effects of MPTP lesioning on the density of DAergic neurons in the substantia nigra pars compacta (SNc). (A–D) The large
square in D indicates the area shown in A–C and the small black square inside it indicates the area in which the density of DAergic...

Chao-Yu Hsu, Ching-Sui Hung, Hung-Ming Chang, Wen-Chieh Liao, Shih-Chun Ho, Ying-Jui Ho

Ceftriaxone prevents and reverses behavioral and neuronal deficits in an MPTP-induced animal model of Parkinson's
disease dementia
Neuropharmacology, Volume 91, 2015, 43–56

http://dx.doi.org/10.1016/j.neuropharm.2014.11.023
 Summary