title: Effect of different media additives on capacitation of frozen-thawed ram spermatozoa as a potential replacement for estrous sheep serum.
abstract: Capacitation is a key process through which spermatozoa acquire their fertilizing ability. This event is required for the successful application of assisted reproductive technologies such as IVF. The aim of the present study was to investigate the effect of using a synthetic oviductal fluid medium supplemented with either heparin-hypotaurine alone, in combination with progesterone (P4), 17β-estradiol (E2), or BSA, or just β-cyclodextrin, in replacement for estrous sheep serum (ESS) for ram sperm capacitation. After incubation in the corresponding media for 15 (time 0) or 60 minutes, sperm function was evaluated by computerized sperm motility analysis and flow cytometry (plasma membrane status and fluidity). Treatments rendering the best results in regards to sperm function parameters related to capacitation were used for an IVF test. Herein, neither heparin-hypotaurine (alone), or in combination with P4, or E2, nor β-cyclodextrin induced capacitation-related changes in frozen-thawed ram spermatozoa. Only the medium supplemented with heparin-hypotaurine-BSA was able to induce changes compatible with in vitro capacitation relating to sperm motility pattern and plasma membrane fluidity, comparable to those in ESS-containing medium. Both media yielded sperm parameter values that differed (P < 0.05) from those obtained in the rest of the media tested. However, after the IVF trial, BSA was unable to support cleavage rates (21.80%) comparable to those obtained with ESS (52.60%; P < 0.05). We conclude that heparin-hypotaurine, P4, E2, β-cyclodextrin, or BSA is not suitable for replacing ESS in capacitation and fertilization media for ram spermatozoa.
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: O
authors.initials: O
authors.lastname: García-Álvarez
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Maroto-Morales
authors.affiliation: Regional Center of Animal Selection and Reproduction (CERSYRA) JCCM, Valdepeñas, Spain.
authors.forename: P
authors.initials: P
authors.lastname: Jiménez-Rabadán
authors.affiliation: Regional Center of Animal Selection and Reproduction (CERSYRA) JCCM, Valdepeñas, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Ramón
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: E
authors.initials: E
authors.lastname: del Olmo
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Iniesta-Cuerda
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Anel-López
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: M R
authors.initials: MR
authors.lastname: Fernández-Santos
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.
authors.forename: J J
authors.initials: JJ
authors.lastname: Garde
authors.affiliation: SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain. Electronic address: anajosefa.soler@uclm.es.
authors.forename: A J
authors.initials: AJ
authors.lastname: Soler
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003470:Culture Media; D004958:Estradiol / Q000494:pharmacology; D005307:Fertilization in Vitro / Q000662:veterinary; D005434:Flow Cytometry; D011374:Progesterone / Q000494:pharmacology; D055101:Semen Analysis / Q000662:veterinary; D012756:Sheep / Q000097:blood / Q000502:physiology*; D013075:Sperm Capacitation / Q000187:drug effects*; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D003470:Culture Media; D047392:beta-Cyclodextrins; D011374:Progesterone; D004958:Estradiol; C031215:betadex
keywords: BSA; Capacitation; Estrous sheep serum; Fertilization; Ram spermatozoon