Up-regulation of cyclin D1 occurs in apoptosis of immature but not mature cerebellar granule neurons in culture
✍ Scribed by Kazuhisa Sakai; Kazuhiko Suzuki; Shuuitsu Tanaka; Tatsuro Koike
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 343 KB
- Volume
- 58
- Category
- Article
- ISSN
- 0360-4012
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✦ Synopsis
Cerebellar granule neurons isolated from 7-day-old rats and cultured in normal medium undergo apoptosis, but remain healthy under depolarizing conditions with elevated K ؉ (м25 mM) or in the presence of brain-derived neurotrophic factor. Northern blot analysis showed that cyclin D1 mRNA was upregulated in this apoptotic process. Both granule neurons and microglia were immunostained with anti-cyclin D1 antibodies, which is consistent with our previous finding that microglia become activated in response to neuronal cell death under these conditions. Only granule neurons, however, showed an enhanced expression of both mRNA and protein levels of cyclin D1 in the presence of aphidicolin that completely eliminated non-neuronal cells. The entire cell body of granule neurons became immunostained prior to cell shrinkage or nuclear condensation. Moreover, cell cycle blockers and an inhibitor of cyclindependent kinases suppressed both increased immunoreactivity and cell death, further substantiating the involvement of an abortive cell cycle in this process. In contrast, both levels of cyclin D1 remained unaltered in mature granule neurons undergoing apoptosis following combined serum withdrawal and low K ؉ shift, suggesting developmental stage dependence of granule neuron apoptosis in vitro. This culture system is suitable for further analysis of the role of cyclin D1 in cell death.