PNS I 2.26.

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Neurons are nondividing cells, and the rules of cell cycle differ with neurons (turning on cell
cycle genes would kill them). After neurogenesis, neurons can migrate to different areas in the
brain via specialized tracks such as radial glia or pioneering axons (different kinds of migration).
After reaching their specific area, they can then differentiate into specific neuron types. After
differentiating, they release axons and dendrites to start communicating and form synapses.
We then have synaptic rearrangement, with synapses forming and pruning (in kids and, less
frequently, in adults).

Starting with the earliest experiments by Spemann in 1924, they took a blastopore and clipped
off a piece at the pore and pasted it onto a different embryo. They got a neural axis that
shouldn’t have been there – essentially a second nervous system, suggesting that the
blastopore lip was an “organizer.” This ectodermal layer on the embryo was defaulted to
become neural tissue, while the other end of this same embryo was producing different factors
(BMP) to differentiate into something else. The reason that one part differentiated into neural
cells but not BMP-influenced was because of inhibitors that prevented the influence of BMP.
These inhibitors at the Spemann site included chordin and noggin. The Spemann organizer is
ubiquitous among organisms, ranging from frogs to zebrafish to mice, and the same region is
present to allow the formation of the nervous system with the same mechanism.

Initially, as the neural tube is forming, the mesoderm forms structures called somites. These
somites will differentiate and form different parts such as the muscle and connective tissue.
The somites are controlled by different factors as to the migration and differentiation. At the
cranial end, the neural tube will balloon out and form the different cephalic parts. Initially, the
embryo will develop very symmetrically, but will “roll over” onto one side and initiate
asymmetric development. Nerve growth factor regulates this process.

As the neural fold is starting to invaginate, there are cells that produce Wnt, which helps
formation of the neural tube and differentiation of neural crest cells. Wnt turns on a bunch of
transcription factors, which then turn on other transcriptions factors (Snail) to block cell death.
C-myc is also turned on (a strong driver of proliferation), and eventually you start turning on
genes that allow neural crest cells to migrate. A lot of this process happens to be analogous to
those in tumor formation. A receptor called Eph (ephrin) is turned on and is involved with
guiding neurons. As these cells are moving (image a sheet of cells moving and folding), known
as planar polarity, the receptor for Wnt, known as Frizzled, is at the front. At the back is a high
concentration of Wnt inhibitor. When Wnt binds to Frizzled, it frees up beta-catenin to move to
the nucleus and regulate transcription. In the case of planar cell polarity, there are small GTP-
binding proteins that are activated by Wnt. So in terms of movement, Wnt regulates the
cytoskeleton, and as it happens, the gradient is at the leading edge of the cell, and at the back
end of the cell, there is a protein called Vangl2 which shuts down Wnt.

Shh is a ventralizing factor and is opposite of BMP, which is a dorsalizing factor. Motor neurons
are induced by Shh, and causes formation of the floor plate. There is therefore a gradient of
BMP and Shh across the neural tube, forming different types of neurons and cells. BMPs bind to
surface receptors and induce certain signals. Type II receptor is always on, but does not
associate with Type I until BMP is bound. Type II will phosphorylate Type I, and phosphorylate
downstream Smads to be then moved to the nucleus to start transcription. In the ventral part
of the neural tube, motor neurons will start forming. Once motor neurons are formed, they will
migrate out and be myelinated. Principle of neuronal formation: neuron, then glia, then
astrocytes. Wnt and BMP are found at the dorsal end, with Shh at the ventral side.

At the same time as these dorsal-ventral factors, there are anterior-posterior factors. There are
inhibitors and activators, noggin/chordin and retinoic acid (a very potent morphogen), at front-
back gradients. One of these things that’s produced in response to retinoic acid is the
rhombomere at the hindbrain. That’s going to lead the formation of some cranial nerve
structures (like the vagus nerve). Some of the cranial sensory neurons are formed from
placodes. During the formation of the neural tube, there are cells on the edge of the ectoderm,
which then form the placode. The placodes form sensory neurons and the lens, portions of the
olfactory system.

Rhombomeres give rise to specific groups of neurons, but what was noticed in expression of
certain genes was that the Hox genes give boundaries of rhombomeres. Bithorax was a
phenotype that was the result of the Hox gene modification (homeobox gene). The gene that
produces the protein is called Hox gene, while the protein itself is the homeobox protein. These
hox genes, which are organized in a way that correlates to their position in the fly (anterior-
posterior to 3’-5’direction). These genes are temporally and spatially segregated. The bithorax
complex was therefore genetically duplicated and phenotypically duplicated as well. These
genes are also induced by retinoic acid and can turn on downstream genes as well. We know
now that this is conserved in vertebrates as well, so that they are turned on in certain regions in
the mouse and in the same temporal sequence.

Now we move to the classic neural crest cells: the rhombomeres form sensory neurons at the
spinal cord. The entire PNS is formed from the neural crest cells as they migrate from the
somites. The migration is regulated by Eph, as the neural crest cells begin to migrate. In the
somite, on one specific part, they express Ephrin (inhibitory) and keeps the migrating cells to
one site to form specific structures. These neural crest cells migrate from dorsal to ventral
sides, as well as anterior to posterior. Sox10 is a marker for these neural crest cells, which is a
precursor for neurons or glia. BMPs will turn on genes in these neural precursors to form
sympathetic and parasympathetic genes. Tyrosine hydroxylase is the rate limiting enzyme in
catecholamine biosynthesis.