title: Sperm Release From the Oviductal Epithelium Depends on Ca(2+) Influx Upon Activation of CB1 and TRPV1 by Anandamide.
abstract: The oviduct acts as a functional sperm reservoir in many mammalian species. Both binding and release of spermatozoa from the oviductal epithelium are mainly modulated by sperm capacitation. Several molecules from oviductal fluid are involved in the regulation of sperm function. Anandamide is a lipid mediator involved in reproductive physiology. Previously, we demonstrated that anandamide, through activation of the cannabinoid receptor type 1 (CB1), promotes sperm release from bovine oviductal epithelial cells, and through CB1 and the transient receptor potential vanilloid 1 (TRPV1), induces sperm capacitation. Herein we investigate co-activation between CB1 and TRPV1, and Ca(2+) influx as part of the mechanism of action of anandamide during sperm release from oviductal cells. Our results indicate that in the absence of Ca(2+) anandamide failed to release spermatozoa from oviductal epithelial cells. Additionally, sperm release promoted by cannabinoid and vanilloid agonists was abolished when the spermatozoa were preloaded with BAPTA-AM, a Ca(2+) chelator. We also determined Ca(2+) levels in spermatozoa preloaded with FURA2-AM co-cultured with oviductal cells and incubated with different cannabinoid and vanilloid agonists. The incubation with different agonists induced Ca(2+) influx, which was abolished by CB1 or TRPV1 antagonists. Our results also suggest that a phospholypase C (PLC) might mediate the activation of CB1 and TRPV1 in sperm release from the bovine oviduct. Therefore, our findings indicate that anandamide, through CB1 and TRPV1 activation, is involved in sperm release from the oviductal reservoir. An increase of sperm Ca(2+) levels and the PLC activation might be involved in anandamide signaling pathway.
authors.affiliation: Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts.|Laboratorio de Biología de la Reproducción en Mamíferos, Centro de Estudios Farmacológicos y Botánicos, (CONICET-UBA), Buenos Aires-Argentina.
authors.forename: M G
authors.initials: MG
authors.lastname: Gervasi
authors.affiliation: Laboratorio de Biología de la Reproducción en Mamíferos, Centro de Estudios Farmacológicos y Botánicos, (CONICET-UBA), Buenos Aires-Argentina.
authors.forename: C
authors.initials: C
authors.lastname: Osycka-Salut
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: T
authors.initials: T
authors.lastname: Sanchez
authors.affiliation: Laboratorio de Biología de la Reproducción en Mamíferos, Centro de Estudios Farmacológicos y Botánicos, (CONICET-UBA), Buenos Aires-Argentina.
authors.forename: C A I
authors.initials: CA
authors.lastname: Alonso
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: C
authors.initials: C
authors.lastname: Llados
authors.affiliation: Laboratorio de Biología de la Reproducción en Mamíferos, Centro de Estudios Farmacológicos y Botánicos, (CONICET-UBA), Buenos Aires-Argentina.
authors.forename: L
authors.initials: L
authors.lastname: Castellano
authors.affiliation: Laboratorio de Fisiopatología de la Preñez y el Parto, Centro de Estudios Farmacológicos y Botánicos (CONICET-UBA), Buenos Aires-Argentina.
authors.forename: A M
authors.initials: AM
authors.lastname: Franchi
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: M
authors.initials: M
authors.lastname: Villalón
authors.affiliation: Laboratorio de Biología de la Reproducción en Mamíferos, Centro de Estudios Farmacológicos y Botánicos, (CONICET-UBA), Buenos Aires-Argentina.
authors.forename: S
authors.initials: S
authors.lastname: Perez-Martinez
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology*; D020013:Calcium Signaling; D063386:Cannabinoid Receptor Agonists / Q000494:pharmacology*; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques; D063388:Endocannabinoids / Q000494:pharmacology*; D005260:Female; D008297:Male; D010057:Oviducts / Q000166:cytology / Q000378:metabolism*; D053284:Polyunsaturated Alkamides / Q000494:pharmacology*; D043884:Receptor, Cannabinoid, CB1 / Q000378:metabolism*; D013075:Sperm Capacitation; D050916:TRPV Cation Channels / Q000378:metabolism*
chemical_list: D001095:Arachidonic Acids; D063386:Cannabinoid Receptor Agonists; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; D043884:Receptor, Cannabinoid, CB1; D050916:TRPV Cation Channels; C078814:anandamide
keywords: BOVINE; CANNABINOID RECEPTORS; Ca2+; SPERM-OVIDUCT INTERACTION; TRPV1