RESUMEN
There is pharmacological evidence that Ca2+ channels play an essential role in triggering the mammalian sperm acrosome reaction, an exocytotic process required for sperm to fertilize the egg. Spermatozoa are small terminally differentiated cells that are difficult to study by conventional electrophysiological techniques. To identify the members of the voltage-dependent Ca2+ channel family possibly present in sperm, we have looked for the expression of the alpha 1A, alpha 1B, alpha 1C, alpha 1D and alpha 1E genes in mouse testis and in purified spermatogenic cell populations with RT-PCR. Our results indicate that all 5 genes are expressed in mouse testis, and in contrast only alpha 1E, and to a minor extent alpha 1A, are expressed in spermatogenic cells. In agreement with these findings, only T-type Ca2+ channels sensitive to the dihydropyridine nifedipine were observed in patch-clamp recordings of pachytene spermatocytes. These results suggest that low-threshold Ca2+ channels are the dihydropyridine-sensitive channels involved in the sperm acrosome reaction.
Asunto(s)
Acrosoma/metabolismo , Canales de Calcio/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo , Animales , Secuencia de Bases , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio/genética , Cartilla de ADN , Dihidropiridinas/farmacología , Expresión Génica , Masculino , Ratones , Datos de Secuencia Molecular , Técnicas de Placa-Clamp , Reacción en Cadena de la Polimerasa , Espermatozoides/efectos de los fármacos , Testículo/citologíaRESUMEN
Ion fluxes through poorly understood channel-mediated mechanisms participate in the interaction between spermatozoa and egg. Previously, we reported the characterization in planar bilayers of a high conductance Ca(2+)-selective, voltage-dependent multistate channel from S. purpuratus sea urchin sperm plasma membranes. Here we show that this ion channel can be directly transferred to planar lipid bilayers upon sperm addition, from sea urchin (S. purpuratus and L. pictus) and from mouse. We found that spermatozoa from these species possess a conspicuous Ca(2+)-selective, high conductance, multi-state, voltage-dependent channel, which displays similar voltage dependence and equal PBa2+/PK+ approximately 4 in the three species. The presence of this Ca2+ channel in such diverse species suggests it plays a relevant role in sperm physiology. The high sensitivity of planar bilayers to detect single ion channels can now be used to study ion channel regulation and gamete interaction.