The expression of calpain 1 and calpain 2 in spermatogenic cells and spermatozoa of the mouse.
There is some evidence suggesting that Ca2+ is involved in processes that occur during the development and function of spermatozoa. Calcium-dependent proteins, such as calmodulin, are expressed during mammalian spermatogenesis further suggesting that Ca2+ takes part in its regulation. However, the precise roles of Ca2+ in spermatogenesis remain to be elucidated. Calpains are a family of Ca(2+)-dependent cysteine proteases whose members are expressed ubiquitously or in a tissue-specific manner. Calpain has been demonstrated to mediate specific Ca(2+)-dependent processes including cell fusion, mitosis and meiosis. We herein followed the expression pattern of calpain's ubiquitous isoforms, 1 and 2, throughout spermatogenesis at the RNA and protein levels by RT-PCR and Western blotting analysis. Both RNA and protein studies revealed that these isoforms are expressed in all spermatogenic cells. The expression of calpain 1 levels is slightly higher in spermatocytes entering the meiotic phase. Both calpain isoforms are also expressed in mouse spermatozoa and are localized to the acrosomal cap. Inducing capacitated spermatozoa to undergo the acrosome reaction in the presence of a selective calpain inhibitor significantly reduced the acrosome reaction rate in a dose-dependent manner. Thus, calpain, a pluripotential protease with numerous substrates, may serve as an effector in more than one pathway in the complex process of spermatogenesis and in the events preceding fertilization, such as the acrosome reaction.
Ben-Aharon
Irit
I
Department of Cell and Developmental Biology, Sackler School of Medicine, Tel-Aviv University, Tel-Aviv, Israel.
Brown
Paula R
PR
Etkovitz
Nir
N
Eddy
Edward M
EM
Shalgi
Ruth
R
D020101:Acrosome Reaction / Q000187:drug effects; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002154:Calpain / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000235:genetics; D004151:Dipeptides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D012313:RNA / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013090:Spermatocytes / Q000737:chemistry; D013091:Spermatogenesis / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry*
D004151:Dipeptides; C071482:calpeptin; D012313:RNA; D002154:Calpain; C112682:Capn1 protein, mouse