title: Anandamide induces sperm release from oviductal epithelia through nitric oxide pathway in bovines.
abstract: Mammalian spermatozoa are not able to fertilize an egg immediately upon ejaculation. They acquire this ability during their transit through the female genital tract in a process known as capacitation. The mammalian oviduct acts as a functional sperm reservoir providing a suitable environment that allows the maintenance of sperm fertilization competence until ovulation occurs. After ovulation, spermatozoa are gradually released from the oviductal reservoir in the caudal isthmus and ascend to the site of fertilization. Capacitating-related changes in sperm plasma membrane seem to be responsible for sperm release from oviductal epithelium. Anandamide is a lipid mediator that participates in the regulation of several female and male reproductive functions. Previously we have demonstrated that anandamide was capable to release spermatozoa from oviductal epithelia by induction of sperm capacitation in bovines. In the present work we studied whether anandamide might exert its effect by activating the nitric oxide (NO) pathway since this molecule has been described as a capacitating agent in spermatozoa from different species. First, we demonstrated that 1 µM NOC-18, a NO donor, and 10 mM L-Arginine, NO synthase substrate, induced the release of spermatozoa from the oviductal epithelia. Then, we observed that the anandamide effect on sperm oviduct interaction was reversed by the addition of 1 µM L-NAME, a NO synthase inhibitor, or 30 µg/ml Hemoglobin, a NO scavenger. We also demonstrated that the induction of bull sperm capacitation by nanomolar concentrations of R(+)-methanandamide or anandamide was inhibited by adding L-NAME or Hemoglobin. To study whether anandamide is able to produce NO, we measured this compound in both sperm and oviductal cells. We observed that anandamide increased the levels of NO in spermatozoa, but not in oviductal cells. These findings suggest that anandamide regulates the sperm release from oviductal epithelia probably by activating the NO pathway during sperm capacitation.
authors.affiliation: Centro de Estudios Farmacológicos y Botánicos (CEFYBO-CONICET), Buenos Aires, Argentina.
authors.forename: Claudia
authors.initials: C
authors.lastname: Osycka-Salut
authors.forename: María Gracia
authors.initials: MG
authors.lastname: Gervasi
authors.forename: Elba
authors.initials: E
authors.lastname: Pereyra
authors.forename: Maximiliano
authors.initials: M
authors.lastname: Cella
authors.forename: María Laura
authors.initials: ML
authors.lastname: Ribeiro
authors.forename: Ana María
authors.initials: AM
authors.lastname: Franchi
authors.forename: Silvina
authors.initials: S
authors.lastname: Perez-Martinez
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology*; D002417:Cattle; D002450:Cell Communication / Q000187:drug effects; D063388:Endocannabinoids; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005187:Fallopian Tubes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D006454:Hemoglobins / Q000494:pharmacology; D008297:Male; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D053284:Polyunsaturated Alkamides / Q000494:pharmacology*; D043884:Receptor, Cannabinoid, CB1 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013094:Spermatozoa / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D050916:TRPV Cation Channels / Q000378:metabolism
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D006454:Hemoglobins; D020030:Nitric Oxide Donors; D053284:Polyunsaturated Alkamides; D043884:Receptor, Cannabinoid, CB1; D050916:TRPV Cation Channels; C088155:methanandamide; D009569:Nitric Oxide; D052248:Nitric Oxide Synthase Type I; D052250:Nitric Oxide Synthase Type III; C078814:anandamide; D019331:NG-Nitroarginine Methyl Ester