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Journal of Neuroscience, Vol 14, 6571-6584, Copyright © 1994 by Society for Neuroscience
Neurotrophin-3 induces neural crest-derived cells from fetal rat gut to develop in vitro as neurons or glia
A Chalazonitis, TP Rothman, J Chen, F Lamballe, M Barbacid and MD Gershon
Department of Anatomy and Cell Biology, Columbia University, College of Physicians and Surgeons, New York, New York 10032.
The precursor cells that form the enteric nervous system (ENS) are
multipotent when they arrive in the gut from the neural crest. Their
differentiation thus depends on signals from the enteric microenvironment.
Crest-derived cells were isolated from the fetal rat bowel by
immunoselection at E14 with NC-1/HNK-1 antibodies and secondary antibodies
coupled to magnetic beads. NC-1/HNK-1- immunoreactive cells were enriched
approximately 36-fold. The NC-1/HNK- 1-selected population and the residual
population were plated at equal cell density and maintained in a defined
medium for 6-7 d. The total number of cells found in the cultures of the
residual cells was three- to fourfold that in cultures of immunoselected
cells. Neurotrophin-3 (NT-3), but not nerve growth factor (NGF),
brain-derived neurotrophic factor (BDNF), or neurotrophin-4/5 (NT-4/5), was
found to increase the proportion of neurons (neurofilament-immunoreactive
or neuron-specific enolase-immunoreactive) or glia (S-100-immunoreactive)
(from 6.6 +/- 0.9% to 15.2 +/- 1.4%; p < 0.001). This effect was
concentration dependent (from 1 to 40 ng/ml) and observed only in the
cultures of immunoselected cells. NT-3 also enhanced neurite outgrowth.
NT-3 increased neither cell number nor bromodeoxyuridine incorporation and
thus was not mitogenic. Exposure of immunoselected cells to NT-3 rapidly
and transiently induced the appearance of nuclear Fos immunoreactivity.
Transcripts coding for TrkC, the transducing receptor for NT-3, were
identified in the fetal rat gut (E14-E16) and in the immunoselected
population of cells using reverse transcriptase and the polymerase chain
reaction. It is concluded that NT-3 specifically promotes the
differentiation of enteric crest-derived cells as neurons or glia and may
thus play a role in the development and/or maintenance of the ENS.
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