|Budget Amount *help
¥8,100,000 (Direct Cost : ¥8,100,000)
Fiscal Year 1996 : ¥2,600,000 (Direct Cost : ¥2,600,000)
Fiscal Year 1995 : ¥5,500,000 (Direct Cost : ¥5,500,000)
Changes in shapes of cells during cell division and cell differentiation are mainly dependent on actin-related cytoskeletons on the cell surface. Calpain, calcium-dependent neutral protease, has proteolytic activity which irreveribly changes the components of the cytoskeleton. In this study, the physiological functions of calpain have been analyzed by developmental and biochmical methods during early embryogenesis of Drosophila melanogaster. First, localization of calpain and actin was analyzed by immunohistochmical methods. Second, cDNA clones for actin-binding proteins, probable substrates for calpain, were obtained and expressed in Escherichia coli together with the expression of active calpain. Third, the probes for calpain reaction, specific antibodies for cleavage sites generated by calpain reaction, were prepared. Fourth, using these probes described above, dynamism of localization of calpain and its substrates were analyzed during Drosophila development. Consequently, the results were obtained that calpain is involved in the formation and reorganization of actin-related cytoskeleton in pseudo-cleavage furrow.