Metabolic and genetic analyses of apoptosis in potassium/serum-deprived rat cerebellar granule cells

Cerebellar granule cells maintained in medium containing serum and 25 mM potassium undergo an apoptotic death within 96 hr when switched to serum- free medium with 5 mM potassium. Because large numbers of apparently homogeneous neurons can be obtained, this represents a potentially useful model of neuronal programmed cell death (PCD). Analysis of the time course and extent of death after removal of either serum or K alone demonstrated that a fast-dying (T( 1/4 ) = 4 hr) population (20%) responded to serum deprivation, whereas a slow-dying (T( 1/4 ) = 25 hr) population (80%) died in response to K⁺ deprivation. Taking advantage of the complete death after removing both K⁺ and serum, changes in metabolic events and mRNA levels were analyzed in this model. Glucose uptake, protein synthesis, and RNA synthesis fell to <35% of control by 9 hr after potassium/serum deprivation, a time when 85% of the cells were still viable. The pattern of the fall in these metabolic parameters was similar to that reported for trophic factor-deprived sympathetic neurons. Most mRNAs decreased markedly after K⁺/serum deprivation. Levels of c-jun mRNA increased fivefold in potassium/serum- deprived granule cells; c-jun is required for cell death of sympathetic neurons. mRNA levels of cyclin D1, c-myb, collagenase, and transin remained relatively constant in potassium/serum-deprived granule cells. These data demonstrate the existence of two populations of granule cells with respect to cell death and define common metabolic and genetic events involved in neuronal PCD.