Changes in calpain during meiosis in the rat egg.
Resumption of meiosis at fertilization is mediated by increased levels of calcium which activate several calcium-dependent enzymes. Calpain, a neutral calcium-activated thiol protease, is present in the cytoplasm of many cells. Its activation is associated with limited autolysis and relocalization in the cell. Calpain is thought to participate in the regulation of mitosis and resumption of meiosis in Xenopus oocytes. In this study we followed the activation and localization of calpain during maturation and fertilization in rat eggs using a polyclonal antibody raised against chicken muscle calpain. A band of 80 kDa was detected in GV oocytes and its level increased in unfertilized MII eggs. At the early stages of fertilization, we observed a transient decrease in the level of calpain which was regained at the pronuclear stage. Adding Ca2+ to lysate of MII eggs resulted in an additional band, representing the degraded fragment of the activated protein. In eggs activated by ionomycin, calpain level decreased, followed by an increase in a dynamic similar to that observed in fertilized eggs. Egg activation also led to changes in calpain localization. A homogenous distribution was observed in GV and in MII eggs, while in activated eggs it was localized predominantly overlying the metaphase plate. In the current study we demonstrate the presence of calpain in the rat egg. During maturation, calpain level increases; however, during egg activation, in response to [Ca2+]i changes, calpain undergoes autolysis, translocaton, and fluctuation in its level. We therefore suggest a correlation between calpain activation and fertilization.
Malcov
M
M
Department of Embryology, Sackler School of Medicine, Tel Aviv University, Israel.
Ben-Yosef
D
D
Glaser
T
T
Shalgi
R
R
D000818:Animals; D002154:Calpain / Q000378:metabolism*; D002645:Chickens; D015151:Immunoblotting; D008540:Meiosis; D010063:Ovum / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D002154:Calpain