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Journal of Neuroscience, Vol 7, 2203-2213, Copyright © 1987 by Society for Neuroscience
The role of depolarization in the survival and differentiation of cerebellar granule cells in culture
V Gallo, A Kingsbury, R Balazs and OS Jorgensen
Cultures greatly enriched in granule cells from early postnatal cerebellum
(P8) were grown in a medium containing fetal calf serum. Under the
conditions used, nerve cells died, usually within a week, unless the K+
concentration in the medium was greater than or equal to 20 mM. The
requirement for elevated [K+]e was manifested by about 3 d in vitro, and
after this time continuous exposure to high [K+]e was essential for the
survival of the granule cells. The initial morphological and biochemical
maturation of the granule cells was similar in the presence and the absence
of elevated [K+]e, suggesting that the dependence on depolarizing
conditions develops in parallel with the expression of the differentiated
characteristics of the cells. The positive effect of elevated [K+]e on
granule cell survival was not influenced by preventing bioelectric activity
in the cultures with TTX and xylocaine. On the other hand,
depolarization-induced transmembrane Ca2+ flux was essential in securing
the maintenance of the granule cells. Depolarized nerve cells were
compromised when Ca2+ entry was blocked by elevated Mg2+, EGTA, or organic
Ca2+ antagonists, while dihydropyridine Ca2+ agonists [BAY K 8644,
(+)-(S)-202 79 1 and CGP 28392] were potent agents preventing nerve cell
loss in the presence of 15 mM [K+]e, which was ineffective on its own.
Calmodulin inhibitors (1 microM trifluoperazine or calmidazolium) blocked
the beneficial effect of K+-induced depolarization on granule cells. The
comparison of the timing of the differentiation and innervation of the
postmitotic granule cells in vivo with the development of the K+ dependence
in vitro would indicate that depolarization of the granule neurons in
culture mimics the influence of the physiological stimulation in vivo
through excitatory amino acid receptors, including N-methyl-D-aspartate
receptors, involving Ca2+ entry and the activation of a Ca2+/calmodulin-
dependent protein kinase.
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G. Wu, X. Xie, Z.-H. Lu, and R. W. Ledeen
Cerebellar neurons lacking complex gangliosides degenerate in the presence of depolarizing levels of potassium
PNAS,
December 22, 2000;
(2000)
11523698.
[Abstract]
[Full Text]
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