Correlação das proteínas do plasma seminal com a congelabilidade do sêmen de cães
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Data
2017-02-21
Autores
Camargo, Laíza Sartori de [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Os objetivos deste estudo foram validar a sonda fluorescente (MitoStatus Red) para análise de potencial mitocondrial em espermatozoide de cães por citometria de fluxo, associada a análise de integridade de membrana plasmática e do acrossomo (experimento I). Além disso, foi conduzida análise proteômica (abordagem shotgun) do plasma seminal de cães, sua correlação com a congelabilidade do sêmen e capacidade de ligação das células espermáticas em membrana perivitelina da gema de ovo (TL) (experimento II). No experimento I foram utilizado 10 ejaculados, com motilidade espermática >75% e as concentrações de 20 nM, 50 nM, 100 nM e 200 nM da sonda MitoStatus Red foram comparadas na citometria de fluxo. Concluiu-se que 20 nM foi a concentração ideal, sem danos as estruturas celulares. No experimento II foram utilizados 10 animais, 2 ejaculados/animal (n = 20). Os ejaculados foram avaliados por cinética espermática, citometria de fluxo (potencial mitocondrial, integridade de membrana plasmática e de acrossomo) e TL. A análise proteômica do plasma seminal foi avaliada por espectrometria de massas. Os ejaculados foram divididos em 3 clusters (cluster Low, cluster Medium, cluster Hight) de acordo com os parâmetros espermáticos pré e pós-descongelação. Os grupos foram correlacionados com as proteínas encontradas no plasma seminal. Concluiu-se que existe relação entre presença ou ausência de proteínas do plasma seminal de acordo com a qualidade espermática.
Canine reproduction studies are important because of it closeness with human reproductive physiology, and it´s the main experimental model for wild species. The lack of studies in canine reproduction of stray animals, is an important tool for the alternative birth control methods researches, is another relevant reason to study reproduction in this species. In order to find fertility or infertility markers, seminal plasma and/or sperm proteins have already been the subject of studies in many other species with different approaches and applications. In farm animals, these proteins have been studied in order to elucidate spermatogenesis physiological events, sperm maturation, fertilization and cell changes when subjected to cryopreservation. Thus, those studies have produced results that can be used to identify animals with higher or lower fertility and freezability. In this sense, and in view of the importance of the dog as experimental model, and because seminal plasma proteins can increase the resistence of sperm cell in front of the cryopreservation process is believed that the identification of the freezability markers, in this species, can generate results applicable to other canids contributing to the formation of germplasm banks. In this way, the aim of this study was to correlate the semen quality and sperm cells binding capacity to the perivitelline egg yolk membrane, pre and post-freezing, with the seminal plasma protein analysis using shotgun approach.
Canine reproduction studies are important because of it closeness with human reproductive physiology, and it´s the main experimental model for wild species. The lack of studies in canine reproduction of stray animals, is an important tool for the alternative birth control methods researches, is another relevant reason to study reproduction in this species. In order to find fertility or infertility markers, seminal plasma and/or sperm proteins have already been the subject of studies in many other species with different approaches and applications. In farm animals, these proteins have been studied in order to elucidate spermatogenesis physiological events, sperm maturation, fertilization and cell changes when subjected to cryopreservation. Thus, those studies have produced results that can be used to identify animals with higher or lower fertility and freezability. In this sense, and in view of the importance of the dog as experimental model, and because seminal plasma proteins can increase the resistence of sperm cell in front of the cryopreservation process is believed that the identification of the freezability markers, in this species, can generate results applicable to other canids contributing to the formation of germplasm banks. In this way, the aim of this study was to correlate the semen quality and sperm cells binding capacity to the perivitelline egg yolk membrane, pre and post-freezing, with the seminal plasma protein analysis using shotgun approach.
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Palavras-chave
Canino, Citometria de fluxo, Spermatozoa, Espectometria de massa, Mass spectometry, Flow citometry, Canine, Semen spermatozoa