Apoptosis occurs as a physiologic process in the ovarian life cycle. Stauro
sporine, a protein kinase inhibitor, is reported to induce apoptosis. Here,
we hypothesize that staurosporine will induce apoptosis in human luteinize
d granulosa cells and that mitochondria and the caspase cascade participate
in this pro cess. Luteinized granulosa cells isolated from in vitro fertil
ization patients were treated with staurosporine. Microscopy revealed that
staurosporine treatment resulted in cells exhibiting evidence of apoptosis,
including cell detachment, loss of cell processes, membrane shrinkage, and
formation of apoptotic bodies. In the staurosporine-treated cells, how cyt
ometry and confocal microscopy showed a decrease in the mitochondrial cardi
olipin levels. Western analysis showed cleavage of caspase-9, an initiator
caspase, of caspase-3, an executioner caspase, and of a caspase substrate,
poly-(ADP-ribose)-polymerase (PARP) in staurosporine-treated cells. These d
ata support our hypothesis and that this is the first demonstration of the
involvement of mitochondria and of cleavage of caspases in human luteinized
granulosa cell apoptosis. This may serve as a useful model to delineate th
e mechanism of apoptosis in the ovary, such as corpus luteum regression. (C
) 2000 Academic Press.