Developmental features of rat cerebellar neural cells cultured in a chemically defined medium

Developmental features of rat cerebellar neural cells cultured in a chemically defined medium We studied some aspects of the differentiation of rat cerebellar neural cells obtained from 8‐day postnatal animals and cultured in a serum‐free, chemically defined medium (CDM). The ability of the cells to take up radioactive transmitter amino acids was analyzed autoradiographically. The L‐glutamate analogue 3H‐D‐aspartate was taken up by astroglial cells, but not by granule neurons, even in late cultures (20 days in vitro). This is in agreement with the lack of depolarization‐induced release of 3H‐D‐aspartate previously observed in this type of culture. In contrast, 3H‐(GABA) was scarcely accumulated by glial‐fibrillary‐acidic‐protein (GFAP)‐positive astrocytes, but taken up by glutamate‐decarboxylase‐positive inhibitory interneurons and was released in a Ca2+‐dependent way upon depolarization: 3H‐GABA evoked release progressively increased with time in culture. Interestingly, the expression of the vesicle‐associated protein synapsin I was much reduced in granule cells cultured in CDM as compared to those maintained in the presence of serum. These data would indicate that in CDM the differentiation of granule neurons is not complete, while that of GABAergic neurons is not greatly affected. Whether the diminished differentiation of granule cells must be attributed only to serum deprivation or also to other differences in the composition of the culture medium remains to be established. 3H‐GABA was avidly taken up also by a population of cells which were not recognized by antibodies raised against GFAP, glutamate decarboxylase, and microtubule‐associated protein 2. These cells exhibited a stellate morphology, were stained by the monoclonal antibody A2B5, and have been characterized elsewhere (Levi et al, 1986) as bipotential precursors of oligodendrocytes and of a subpopulation of astrocytes bearing a stellate shape and capable of high‐affinity 3H‐GABA uptake. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neuroscience Research Wiley

Developmental features of rat cerebellar neural cells cultured in a chemically defined medium

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Publisher
Wiley
Copyright
Copyright © 1986 Alan R. Liss, Inc.
ISSN
0360-4012
eISSN
1097-4547
DOI
10.1002/jnr.490150302
pmid
3084804
Publisher site
See Article on Publisher Site

Abstract

We studied some aspects of the differentiation of rat cerebellar neural cells obtained from 8‐day postnatal animals and cultured in a serum‐free, chemically defined medium (CDM). The ability of the cells to take up radioactive transmitter amino acids was analyzed autoradiographically. The L‐glutamate analogue 3H‐D‐aspartate was taken up by astroglial cells, but not by granule neurons, even in late cultures (20 days in vitro). This is in agreement with the lack of depolarization‐induced release of 3H‐D‐aspartate previously observed in this type of culture. In contrast, 3H‐(GABA) was scarcely accumulated by glial‐fibrillary‐acidic‐protein (GFAP)‐positive astrocytes, but taken up by glutamate‐decarboxylase‐positive inhibitory interneurons and was released in a Ca2+‐dependent way upon depolarization: 3H‐GABA evoked release progressively increased with time in culture. Interestingly, the expression of the vesicle‐associated protein synapsin I was much reduced in granule cells cultured in CDM as compared to those maintained in the presence of serum. These data would indicate that in CDM the differentiation of granule neurons is not complete, while that of GABAergic neurons is not greatly affected. Whether the diminished differentiation of granule cells must be attributed only to serum deprivation or also to other differences in the composition of the culture medium remains to be established. 3H‐GABA was avidly taken up also by a population of cells which were not recognized by antibodies raised against GFAP, glutamate decarboxylase, and microtubule‐associated protein 2. These cells exhibited a stellate morphology, were stained by the monoclonal antibody A2B5, and have been characterized elsewhere (Levi et al, 1986) as bipotential precursors of oligodendrocytes and of a subpopulation of astrocytes bearing a stellate shape and capable of high‐affinity 3H‐GABA uptake.

Journal

Journal of Neuroscience ResearchWiley

Published: Jan 1, 1986

References

  • Synapsin I (Protein I), a nerve terminal‐specific phosphoprotein. II. Its specific association with synaptic vesicles demonstrated by immunocytochemistry in agarose‐embedded synaptosomes
    De Camilli, De Camilli; Harris, Harris; Huttner, Huttner; Greengard, Greengard

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