Calcium signalling and the regulation of apoptosis.
Apoptotic cell death is characterized by cell shrinkage, chromatin condensation and fragmentation, formation of apoptotic bodies and phagocytosis (Kerr et al., 1972). At the molecular level, activation of a family of cysteine proteases, caspases, related to interleukin-1beta-converting enzyme is believed to be a crucial event in apoptosis. This is associated with the proteolysis of nuclear and cytoskeletal proteins, cell shrinkage, glutathione efflux, exposure of phosphatidylserine on the cell surface, membrane blebbing, etc. In CD95- or TNF-mediated apoptosis, the proteolytic cascade is believed to be triggered directly by caspase binding to the activated plasma membrane receptor complex. In other forms of apoptosis, the mechanisms of activation of the proteolytic cascade are less well established but may involve imported proteases, such as granzyme B, or factors released from the mitochondria and, possibly, other organelles. Recently, the possibility that cytochrome c released from the mitochondria may serve to activate dormant caspases in the cytosol, and thereby to propagate the apoptotic process, has attracted considerable attention. A perturbation of intracellular Ca(2+) homeostasis has been found to trigger apoptosis in many experimental systems, and the apoptotic process has been related to either a sustained increase in cytosolic free Ca(2+) level or a depletion of intracellular Ca(2+) stores. Although many of the biochemical events involved in the apoptotic process are Ca(2+) dependent, the exact mechanism by which Ca(2+) triggers apoptosis remains unknown. The bcl-2 gene family, which includes both inhibitors and inducers of apoptosis, appears to regulate intracellular Ca(2+) compartmentalization. The induction of apoptosis by Ca(2+)-mobilizing agents results in caspase activation, which is similar to what is seen with other inducers of apoptosis. In addition, Ca(2+)-dependent proteases, such as calpain and a Ca(2+)-dependent nuclear scaffold-associated serine protease, are also activated by Ca(2+) signalling in some cell types where they appear to be involved in alpha-fodrin and lamin beta cleavage, respectively. Thus, a spectrum of proteases are activated during apoptosis depending on both cell type and inducer. This proteolytic cascade can involve both caspases and Ca(2+)-dependent proteases, which seem to interact during the apoptotic process.