Expression and possible involvement of calpain isoforms in mammalian egg activation.
At fertilization in mammals, the spermatozoon triggers a unique signal transduction mechanism within the egg, leading to its activation. It is well accepted that the earliest event observed in all activated eggs is an abrupt rise in intracellular calcium concentrations. However, little is known regarding the downstream proteins that are activated by this rise in calcium. Calpains constitute a family of intracellular calcium-dependent cysteine proteases whose members are expressed widely in a variety of cells. We investigated the expression and possible role of the calpain isoforms mu and m throughout egg activation. Both calpains were expressed in the rat egg and localized at the egg cortex as well as in the meiotic spindle. m Calpain translocated to the membrane and to the spindle area during parthenogenetic egg activation and during in vivo fertilization, upon sperm binding to the egg. The cytoskeletal protein alpha-spectrin (fodrin) was proteolysed by calpain during the egg-activation process, as demonstrated by specific calpain-breakdown products. Following parthenogenetic activation by ionomycin or puromycin, the calpain-selective permeable inhibitor, calpeptin, inhibited the resumption of meiosis and cortical reaction in a dose-dependent manner. Calpeptin was also effective in inhibiting in vitro fertilization. These results may imply a correlation between calpain activation and mammalian egg activation at fertilization and a possible role for calpain in the cascade of cellular events leading to resumption of meiosis.
Ben-Aharon
Irit
I
Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, Tel-Aviv, Israel.
Haim
Karin
K
Shalgi
Ruth
R
Ben-Yosef
Dalit
D
D000818:Animals; D020013:Calcium Signaling / Q000502:physiology*; D002154:Calpain / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004151:Dipeptides / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro; D005455:Fluorescent Antibody Technique; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D008540:Meiosis / Q000187:drug effects; D018613:Microscopy, Confocal; D009842:Oligopeptides / Q000494:pharmacology; D010063:Ovum / Q000737:chemistry / Q000378:metabolism*; D010312:Parthenogenesis; D011691:Puromycin / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013084:Sperm-Ovum Interactions / Q000502:physiology*
D004151:Dipeptides; D007476:Ionophores; D009842:Oligopeptides; C069861:calpain inhibitor 2; C071482:calpeptin; D011691:Puromycin; D015759:Ionomycin; D002154:Calpain; C449000:m-calpain; C449002:mu-calpain