Human spermatozoa possess a calcium-dependent chloride channel that may participate in the acrosomal reaction

摘要

Motility, maturation and the acrosome reaction (AR) are fundamental functions of mammalian spermatozoa. While travelling through the female reproductive tract, spermatozoa must mature through a process named capacitation, so that they can reach the egg and undergo the AR, an exocytotic event necessary to fertilize the egg. Though Cl - is important for sperm capacitation and for the AR, not much is known about the molecular identity of the Cl - transporters involved in these processes. We implemented a modified perforated patch-clamp strategy to obtain whole cell recordings sealing on the head of mature human spermatozoa. Our whole cell recordings revealed the presence of a Ca 2+-dependent Cl - current. The biophysical characteristics of this current and its sensitivity to niflumic acid (NFA) and 4,4′-diisothiocyano-2,2′-stilbene disulphonic acid (DIDIS) are consistent with those displayed by the Ca 2+-dependent Cl - channel from the anoctamin family (TMEM16). Whole cell patch clamp recordings in the cytoplasmic droplet of human spermatozoa corroborated the presence of these currents, which were sensitive to NFA and to a small molecule TMEM16A inhibitor (TMEM16A inh, an aminophenylthiazole). Importantly, the human sperm AR induced by a recombinant human glycoprotein from the zona pellucida, rhZP3, displayed a similar sensitivity to NFA, DIDS and TMEM16A inh as the sperm Ca 2+-dependent Cl - currents. Our findings indicate the presence of Ca 2+-dependent Cl - currents in human spermatozoa, that TMEM16A may contribute to these currents and also that sperm Ca 2+-dependent Cl - currents may participate in the rhZP3-induced AR.