Expression of cyclin A decreases during neuronal apoptosis in cultured rat cerebellar granule neurons

Abstract

Cultured cerebellar granule neurons died in an apoptotic manner when the K+ concentration in culture medium was lowered to the normal level (5 mM) after maturation of cells with a high concentration of K+ (26 mM). The changes in expression of 14 cell cycle-related genes in this CNS apoptosis model were analyzed by quantitative RT-PCR. Most of the genes analyzed were stable during apoptosis. The expression of cyclin A mRNA, however, transiently decreased 1 h after the induction of apoptosis, and recovered within 3 h to above the basal level. In this system, the level of cyclin D1, which has been reported to be up-regulated in apoptosis of NGF-deprived cultured sympathetic neurons, did not change. These results suggest that the molecular mechanisms in these two apoptosis models are different. To determine cyclin A protein level, we used an immunostaining method. The number of cyclin A-positive neurons decreased during apoptosis. Moreover, the numbers of MAP2- and cdk2-positive neurons also decreased in a similar manner. Taken together, these results suggest that there is a relationship between apoptosis and cell cycle, and that morphological changes during apoptosis result from cytoskeletal structure degradation.