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Journal of Neuroscience, Vol 14, 29-38, Copyright © 1994 by Society for Neuroscience
GABA-induced chemokinesis and NGF-induced chemotaxis of embryonic spinal cord neurons
TN Behar, AE Schaffner, CA Colton, R Somogyi, Z Olah, C Lehel and JL Barker
Laboratory of Neurophysiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892.
During CNS development, neuroblasts proliferate within germinal zones of
the neuroepithelium, and then migrate to their final positions. Although
many neurons are thought to migrate along processes of radial glial fibers,
increasing evidence suggests environmental factors also influence nerve
cell movement. Extracellular matrix molecules are thought to be involved in
guiding neuronal migration, and molecules such as NGF and GABA exert
trophic effects on immature neurons. The nature of the signals that
initiate and direct neuroblast migration, however, is unknown. In vitro,
NGF and GABA promote neurite outgrowth from cultured cells, and NGF induces
axonal chemotaxis (directed migration along a chemical gradient). At
earlier developmental stages, these molecules could influence neuroblast
movement. Therefore, we investigated whether these molecules induce
embryonic neuronal migration. Using an in vitro microchemotaxis assay, we
show that rat embryonic spinal cord neurons migrate toward picomolar NGF
and femtomolar GABA beginning at embryonic day 13 (E13). Cells exhibit
chemotactic responses to NGF while GABA stimulates chemokinesis (increased
random movement). GABA effects are mimicked by muscimol and inhibited by
bicuculline and picrotoxin, suggesting GABA motility signals are mediated
by GABA receptor proteins. Expression of GABA receptors by embryonic cord
cells has been previously reported (Mandler et al., 1990; Walton et al.,
1993). We used polymerase chain reaction analysis to demonstrate the
presence of NGF and trk mRNA in E13 and E14 cord cells, indicating the
cells express message for both NGF and high- affinity NGF receptors.
Immunohistochemistry of E13 spinal cord sections indicates that NGF and
GABA colocalize in fibers close to the target destinations of migrating
neurons, suggesting diffusible gradients of these molecules provide
chemoattractant signals to migratory cells. Thus, in vitro, neuroblast
migration is induced by specific signaling molecules that are present in
the developing spinal cord, and may stimulate migration of embryonic
neurons prior to synaptogenesis.
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