Study guide- Neural development 3

Neural crest
 What cells give rise to the neural crest? Where are they found?
The neural crest forms when the neural plate invaginates to form the neural tube. There are cells at the crest
of the invagination that pinch off and form the neural rest, which resides between the neural tube and
epidermis.

 What signals help generate the neural crest cells within the ectoderm?
Neural crest cells are specified by BMP, Wnt, and FGF signaling. These signals specify a subset of cells to
become neural crest cells at the border between the non-neural ectoderm or epidermis and the neural plate.

 What is meant by epithelial-mesenchymal transition? If you don’t know what an epithelium or a mesenchyme is,
you might want to look it up.
Epithelial-mesenchymal transition refers to the process by which epithelial cells lose their cell polarity and cell-
cell adhesion to become migratory mesenchymal stem cells, which can differentiate into a variety of cell
types. In this case, the neural crest cells on the ectoderm lose their epithelial like epidermal qualities and
migrate.

 Is neural crest migration random, or does it follow cues?

Neural crest migration follows cues; they share a lot of similar signaling cues to growth cones including repulsive
cues such as Ephrin B and Semaphorin. Because of this, they prefer to migrate along the rostral portion of
neural tube cells because they are repulsed by the caudal portion.

 How is a chick-quail chimera used to trace the normal fate of neural crest cells? How is it used to test whether
pre-migratory neural crest cells are already fixed in their eventual fate?
Similar to other transplantation studies, you can transplant cells from a quail into a chick, or vise versa, and allow
those cells to migrate and differentiate. You can then distinguish whether if the cell you’re looking at came from
the location of transplantation or from the host by looking at their nuclei in this case, or using an antibody that is
specific for quail or chick cells.

Turns out that a specific region of the neural crest along the spinal cord will typically become cholinergic
neurons, while another region of the neural crest along the spinal cord will typically become adrenergic
neurons. So you can then transplant a neural crest cell from a quail from the region where it will typically
become a cholinergic neuron into a chick embryo where it should become adrenergic, and this neural crest cell
will become adrenergic. This means that the neural crest cells are not fixed in their eventual fate, and relies
on other cues externally to identify their cell fate.

 What is the evidence that pre-migratory crest cells can switch between sympathetic and parasympathetic fates?
What was originally used to indicate which fate the cells chose?
Similar answer to the previous question. Chick-quail chimeras were used to determine that pre-migratory crest
cells can switch between sympathetic and parasympathetic fates. Because at the time it was relatively easy to
determine a cell identity by which neurotransmitter they produced, they determined the fate of neural crest cells
by either their production of norepinephrine (sympathetic) or acetylcholine (parasympathetic).

 Are the fates of all neural crest cells unfixed, or are some fixed before migration?
While most neural crest cells have unfixed cell fates, some are fixed before migration. This is particularly
true for the cranial neural crest cells, which have some fixed fates depending on their anterior-posterior
position prior to migration. This is because based on their position, these neural crest cells express different
levels and combinations of HOX genes, which provides spatial memory and contributes to some cell fate identity.
Radial axis:

 What are the two precursor cell types in the ventricular zone? What is a stem cell?
The two precursor cell types in the ventricular zone are the radial glial cells and the intermediate precursor
cells (IPCs). The radial glial cells produce immature neurons and also the dividing intermediate precursor
cells, both of which are self-renewing. However, the intermediate precursor cells can also divide into post-
mitotic cells that migrate outward along radial glia and give rise to immature neurons and glia.

 What labeling technique is used to birthdate post-mitotic cells in the developing neural tube?
A label that consists of some DNA component is used to birthdate post-mitotic cells in the developing neural
tube, which consists of either radioactive H3-thymidine (a nucleic acid) or BRDU, which labels DNA in the S
phase of the cell cycle. The idea is that when a cell is dividing, it will incorporate these labels into the nucleus.
When a cell stops dividing, it will carry this label at high concentrations throughout the cell’s life; however, if
a cell continues to divide, it will dilute this label and eventually become undetectable. So, during development
you can label dividing cells with these markers at different time points, and once they switch into a post-mitotic
cell you can track their fate as they migrate as the host matures.

 In what layers of the brain do post-mitotic cells wind up if they are born early in development? What if they are
born later in development?
Post-mitotic cells early in development end up in the inner layers first, and as development occurs the post-mitotic
cells migrate to the outer layers.

 How are heterochronic transplantation experiments used to test where and when cells acquire layer-specific
identities? Do cells from a young ventricular zone behave the same as cells from an older ventricular zone in
these transplantation experiments? Are cells acquiring layer-specific identities before or after migration into a
layer?
To test if a single dividing cell in the ventricular zone can divide and give rise to cells of multiple layers or
identities, you can label a single dividing cell in a young ventricular zone with some lineage dye or a non-
reproductive virus expressing some marker gene. You then allow the animal to age and allow these single
cells to divide and see what cell in which layer it gives rise to. A single precursor cell can in fact give rise to
all cell types in many different cell layers; so a young ventricular zone precursor cell does not yet know what cell
type and which layer it will migrate to.

A heterochronic transplantation involves transplanting a cell from one aged animal to another differently aged
animal; i.e. a young animal into an old animal, or an old animal into a young animal. The idea is that a younger
ventricular zone cell should be making the inner most layer cells, and the older ventricular zone cell should
be making outer most layer cells. So, if you take a young ventricular zone cell and transplant it into an older
ventricular zone animal, different things can happen depending on which cell cycle it is in. If you transplant a
young ventricular zone cell into an older ventricular zone where the cells are already post-mitotic, just after
their last S phase, they migrate into the inner layers. Thus, once the ventricular zone becomes post-mitotic, it
has already decided where it will go. However, if you transplant a young ventricular zone cell into an older
ventricular zone animal where the cells are pre-S phase, they then migrate to the outer layers. So there is some
cue during and around the S phase of the cell cycle that regulates a cell identity of a ventricular zone cell.
*However, if you take a old ventricular zone cell and transplant it into a young ventricular zone, it will migrate to
the outer layer; so older cells already know their identitity.