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02627_2 T-TYPE CA 2+ CHANNELS MEDIATE SAMMA-INDUCED HUMAN ACROSOMAL LOSS Anderson, Robert A* Work is ongoing to understand the actions of the contraceptive microbicide, SAMMA, a novel low molecular weight carboxylated oligomer. Past work suggested that SAMMA induces acrosomal loss (AL) in human sperm by activating voltage-dependent Ca2+ channels, though probably not L-type. The present study extended observations with Ca2+ channel antagonists and examined selected Ca2+-dependent pathways. Thapsigargin (THG; releases intracellular Ca2+ stores) also induces AL; 1 µM THG causes a 93% (90% confidence limits = 84.0-98.5%) maximal AL; 10 µM nifedipine inhibits this response by 68%. 2-aminoethoxydiphenylborate (2-APB) prevents inositol triphosphate-mediated release of intracellular Ca2+. Consistent with inhibition by nifedipine of THG-induced AL and its failure to block SAMMA-induced AL, 2-APB is without effect. Diphenylhydantoin, a T-type Ca2+ channel blocker, inhibits SAMMA-induced AL (IC50 - 200 µM). Neither trifluoperazine nor calmodulin binding domain affect SAMMA-induced AL, arguing against a role of Ca 2+ /calmodulin-dependent protein kinase II (CaMKII). The data suggest that Ca2+ entry, via either L- or T-type calcium channels, causes AL. CaMKII, involved in AL caused by sustained increases in intracellular Ca2+ (Fert. Steril. (Progr Suppl): S181, 1992), isnt important in SAMMA-induced AL. The oocyte zona pellucida (natural AL stimulus) may cause Ca2+ entry through L-type channels, activation of which are triggered by release of intracellular Ca2+ stores (Mol. Hum. Reprod. 3: 195, 1997.). Similar signalling may accompany epithelial cell infection by HSV (J Cell Biol 163: 283, 2003). SAMMA may be contraceptive and microbicidal by altering the normal Ca2+ responses to viral and sperm entry into target cells. Supported by NIH grants HD41763 and AI37940. Dr. Robert Anderson |
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