title: Post-thaw addition of seminal plasma reduces tyrosine phosphorylation on the surface of cryopreserved equine sperm, but does not reduce lipid peroxidation.
abstract: The objective was to verify the relationship between equine semen cryopreservation and changes related to increased lipid peroxidation. Also, addition of autologous or homologous seminal plasma from a stallion with a good freezing response to post-thawed sperm was tested to determine whether it would confer protection. Frozen-thawed sperm were evaluated and allocated into three groups: without plasma addition, and supplemented with either homologous or autologous seminal plasma. All groups were evaluated at 0, 60 and 120 min after incubation at 37 °C. Cryopreservation did not increase plasma membrane disorders (mean ± SEM 9.48 ± 0.65 and 1.62 ± 0.23% in raw and frozen-thawed sperm, respectively). However, both membrane peroxidation and protein phosphorylation were increased (P < 0.05) compared to raw semen (1.74 and 5.20-fold, respectively). There was a correlation (r = 0.73; P < 0.05) between the increase in lipid peroxidation and tyrosine phosphorylation. Seminal plasma, regardless of origin, reduced (P > 0.05) tyrosine phosphorylation present on the surface of cryopreserved sperm; however, lipid peroxidation was not significantly reduced. In conclusion, we inferred that emergence of phosphorylated proteins on the surface of cryopreserved sperm was due to increased lipid peroxidation that occurred during the freezing/thawing process; however, reduced tyrosine phosphorylation that occurred after addition of seminal plasma was triggered by other mechanisms, apparently independent from the reduction in membrane peroxidation.
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo (USP), Pirassununga, SP, Brazil.
authors.forename: André Furugen Cesar
authors.initials: AF
authors.lastname: de Andrade
authors.forename: Fabiane Gilli
authors.initials: FG
authors.lastname: Zaffalon
authors.forename: Eneiva Carla Carvalho
authors.initials: EC
authors.lastname: Celeghini
authors.forename: Juliana
authors.initials: J
authors.lastname: Nascimento
authors.forename: Fabiana Fernandes
authors.initials: FF
authors.lastname: Bressan
authors.forename: Simone Maria Massami Kitamura
authors.initials: SM
authors.lastname: Martins
authors.forename: Rubens Paes
authors.initials: RP
authors.lastname: de Arruda
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000662:veterinary*; D005434:Flow Cytometry; D005615:Freezing; D006736:Horses / Q000502:physiology*; D015227:Lipid Peroxidation / Q000502:physiology*; D008297:Male; D010766:Phosphorylation; D012661:Semen / Q000502:physiology*; D012662:Semen Preservation / Q000662:veterinary*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D014443:Tyrosine