Recent results have demonstrated that substantial calcium influx in rat Sertoli cells is mediated by cation channels of both L- and N-type. In this report we have investigated the possible role of such channels in the protein secretion of immature rat Sertoli cell monolayers. The blocking of N-type voltage-gated channels by omega-conotoxin (omega-CTX) GVIA results in a 50-60% inhibition of the protein secretion in the culture medium while total protein and RNA synthesis are not affected. The same extent of protein secretion inhibition is obtained in FSH-stimulated Sertoli cells. L-type voltage-gated channels apparently are not involved in such a modulation. These data, showing that a major fraction of secreted proteins from cultured rat Sertoli cells is Ca2+ dependent, represent the first evidence of a physiological role of voltage-operated Ca2+ channels in mammalian testis.