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Journal of Neuroscience, Vol 15, 1172-1179, Copyright © 1995 by Society for Neuroscience
Apoptosis in cerebellar granule cells is blocked by high KCl, forskolin, and IGF-1 through distinct mechanisms of action: the involvement of intracellular calcium and RNA synthesis
C Galli, O Meucci, A Scorziello, TM Werge, P Calissano and G Schettini
Institute of Neurobiology-CNR, Rome, Italy.
Cerebellar granule cells deprived of depolarizing concentration of
extracellular potassium, [K+]o, undergo apoptosis. We here report that this
apoptotic process is associated with an immediate and permanent decrease in
the levels of free intracellular calcium, [Ca2+]i. Although forskolin and
IGF-1 are both able to prevent apoptosis, only forskolin is able to
counteract the instantaneous decrease of [Ca2+]i. However, the early effect
of forskolin on [Ca2+]i is lost after longer incubation in low [K+]o. The
calcium antagonist nifedipine is able to inhibit the survival effect of
high [K+]o, while not affecting forskolin and IGF-1 promoted survival, as
assessed by viability and genomic DNA analysis. Accordingly, the L-type
calcium channels agonist Bay K8644 significantly enhanced the survival of
low KCl treated neurons. To temporally characterize the signal transduction
events and the essential transcriptional step in cerebellar granule cells
apoptosis, we determined the time course of the rescue capacity of high
[K+]o, forskolin, IGF-1, and actinomycin D. Addition of high KCl,
forskolin, or IGF-1 6 hr after the initial KCl deprivation saves 50% of
cells. Remarkably, 50% of neurons loss the potential to be rescued by
actinomycin D after only 1 hr in low [K+]o. Finally, we show that the
survival promoting activities of high [K+]o, forskolin, and IGF-1 do not
require RNA synthesis. We conclude that [Ca2+]i is involved in the survival
promoting activity exerted by high [K+]o but not in those of forskolin and
IGF-1, and that all three agents, although rescuing neurons from apoptosis
through distinct mechanisms of action, do not necessitate RNA
transcription.
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