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Journal of Neuroscience, Vol 10, 1276-1285, Copyright © 1990 by Society for Neuroscience
Neuron-glia interactions of rat hippocampal cells in vitro: glial- guided neuronal migration and neuronal regulation of glial differentiation
UE Gasser and ME Hatten
Department of Pathology, College of Physicians & Surgeons of Columbia University, New York, New York 10032.
To examine neuron-glia interactions of hippocampal cells, including
glial-guided neuronal migration, glial organization of neuronal positioning
and neuronal regulation of astroglial differentiation, rat hippocampal
tissue, harvested between embryonic day 16 (E16) and postnatal day 3 (P3),
was dissociated into a single cell suspension and plated in glass coverslip
microcultures (Hatten and Liem, 1981; Hatten et al., 1984). Immunostaining
the cells with antibodies against the glial filament protein (AbGFP)
revealed developmental stage-specific changes in the number and extent of
morphological differentiation of hippocampal astroglial cells. At E16-E18,
fewer than 5% of the cells were AbGFP-positive; stained cells were
immature, bearing very short processes. By E19-E20, the number of stained
cells increased to 15% of the total cell population. Three forms of
differentiated glial cells predominated, a bipolar form bearing processes
30-50 microns, an elongated form which resembled the radial glia of
hippocampus, bearing processes 120 microns in length, and a stellate form
with 3 or more processes 30-50 microns in length. At P0-P3, glial
morphological differentiation varied with the culture substratum;
differentiated forms resembling those seen at E20 occurred on Matrigel, but
not on polylysine. Quantitation of the distribution of neurons relative to
AbGFP-stained glial processes revealed developmental stage-specific changes
in glial organization of neuronal positioning in the cultures. In cultures
of E16-E18 hippocampal cells, the neurons did not preferentially associate
with astroglial cells. By E19-E20, extensive neuron-glia interactions
occurred, with 80-90% of the neurons being located within 5-10 microns of a
glial process. In addition to their organization of neuronal positioning,
E20 hippocampal astroglial cells supported extensive neuronal migration.
Migrating hippocampal neurons displayed a cytology and neuron-glia cell
apposition identical to that described for migrating cerebellar granule
cells in vitro (Edmondson and Hatten, 1987), closely apposing their cell
soma against the hippocampal glial process and moving along the glial arm
by extending a thickened, leading process. Migration was seen only along
highly elongated glial profiles resembling radial glial seen in vivo. The
morphological differentiation of hippocampal glial cells in vitro was
dependent on cell-cell interactions with neurons. In the absence of
neurons, purified hippocampal astroglia had flat, undifferentiated profiles
and proliferated rapidly. The addition of hippocampal neurons rapidly
arrested glial growth and induced glial process extension.
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