title: Human spermatozoa possess a calcium-dependent chloride channel that may participate in the acrosomal reaction.
abstract: 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 Ca²⁺-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 Ca²⁺-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 (TMEM16Ainh, 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 TMEM16Ainh as the sperm Ca²⁺-dependent Cl⁻ currents. Our findings indicate the presence of Ca²⁺-dependent Cl⁻ currents in human spermatozoa, that TMEM16A may contribute to these currents and also that sperm Ca²⁺-dependent Cl⁻ currents may participate in the rhZP3-induced AR.
authors.affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, CP 62210, México.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Orta
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Ferreira
authors.forename: Omar
authors.initials: O
authors.lastname: José
authors.forename: Claudia L
authors.initials: CL
authors.lastname: Treviño
authors.forename: Carmen
authors.initials: C
authors.lastname: Beltrán
authors.forename: Alberto
authors.initials: A
authors.lastname: Darszon
mesh_terms: D020101:Acrosome Reaction / Q000502:physiology*; D000075369:Anoctamin-1; D002118:Calcium / Q000502:physiology; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000502:physiology*; D006801:Humans; D008297:Male; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D013094:Spermatozoa / Q000502:physiology*; D013844:Thiazoles / Q000494:pharmacology
chemical_list: C476213:ANO1 protein, human; D000075369:Anoctamin-1; D018118:Chloride Channels; D009363:Neoplasm Proteins; D013844:Thiazoles; D002118:Calcium