Portal de Pesquisa da BVS Veterinária

Tese em Português | VETTESES | ID: vtt-217421

Resumo

O presente estudo teve como objetivo descrever o perfil proteico do plasma seminal de búfalos e bovinos e identificar proteínas que funcionem como marcadores de funções espermáticas. Foram utilizados 16 búfalos e 16 bovinos sadios, de 2,5 a 5 anos de idade. A colheita do sêmen foi realizada por eletroejaculação e analisado segundo características macroscópicas e microscópicas. Após a análise, as amostras foram centrifugadas a 800g durante 10 minutos para a separação do plasma seminal, o qual foi imediatamente congelado. No laboratório, as amostras foram descongeladas em banho de gelo e recentrifugadas a 10.000g durante 30 minutos, a 4° C. A concentração de proteína total foi realizada pelo método Bradford utilizando nanoespectrofotômetro As amostras foarm digeridas in solution, seguida da análise de espectrometria de massas. No primeiro experimento, foram encontradas 48 proteínas no plasma seminal de búfalos, sendo que 10 destas apresentaram diferenças estatísticas significativas entre os grupos. As proteínas spermadhesin, ribonuclease, 14-3-3 protein zeta/delta, acrosin inhibitor, epididymal secretory protein E, serum albumin e clusterin foram encontradas em maior abundância no plasma seminal de búfalos de alta motilidade espermática, sendo as demais prosaposin, peptide YY, cystatn-C e secretoglobin family 1D member encontradas em animais de baixa motilidade espermática. No experimento 2 foram identificados um total de 25 proteínas relevantes em bovinos e búfalos, sendo comuns para ambos os grupos apenas 5: secretoglobin family 1D member, serum albumin, clusterin, spermadhesin e epididymal secretory protein E1. No grupo búfalos foram encontradas 4 proteínas: peptide YY, prosaposin, cystatin C e keratin. Já no grupo bovinos 15 proteínas foram relevantes: osteopontin, nucleobidin, acrosin, ribonuclease 4, seminal plasma protein PDC-109, seminal plasma protein A3, seminal plasma protein BSP-30 kDa, C-type natriuretic peptide, serine protease inhibitor Kazal-type 6, C-C motif chemokine 2, angiogenin, metalloproteinase inhibitor 2, seminal ribonuclease, ephrin-A1 e caltrin. O experimento 1 identificou e correlacionou proteínas do plasma seminal de búfalos com a motilidade espermática do sêmen, sendo estas proteínas possíveis marcadores de qualidade espermática na espécie. O experimento 2 comparou o perfil proteico do plasma seminal de búfalos e bovinos e encontrou diferenças proteicas entre as espécies.

---

The aim of the present study was to describe the protein profile of bovine and buffalo seminal plasma and to identify proteins that function as markers of sperm function. Sixteen buffalo and 16 healthy cattle, 2.5 to 5 years old, were used. The semen collection was performed by electroejaculation and analyzed according to macroscopic and microscopic characteristics. After the analysis, the samples were centrifuged at 800g for 10 minutes for separation of the seminal plasma, which was immediately frozen. In the laboratory, the samples were thawed in an ice bath and recentrifuged at 10,000g for 30 minutes at 4°C. Total protein concentration was performed by the Bradford method using nanospectrophotometer. The samples were digested in solution, followed by analysis of spectrophotometry pastas. In the first experiment, 48 proteins were found in the seminal plasma of buffaloes, 10 of which showed significant statistical differences between the groups. The proteins spermadhesin, ribonuclease, 14-3-3 protein zeta / delta, acrosin inhibitor, epididymal secretory protein E, serum albumin and clusterin were found in greater abundance in seminal plasma of buffaloes with high sperm motility, being the other prosaposin, peptide YY , cystatn-C and secretoglobin family 1D member found in animals of low sperm motility. In the experiment 2, a total of 25 relevant proteins were identified in bovines and buffaloes, with only 5 groups: secretaglobin family 1D member, serum albumin, clusterin, spermadhesin and epididymal secretory protein E1. In the buffalo group were found 4 proteins: peptide YY, prosaposin, cystatin C and keratin. In the bovine group 15 proteins were relevant: osteopontin, nucleobidin, acrosin, ribonuclease 4, seminal plasma protein PDC-109, seminal plasma protein A3, seminal plasma protein BSP-30 kDa, C-type natriuretic peptide, serine protease inhibitor Kazal-type 6, CC motif chemokine 2, angiogenin, metalloproteinase inhibitor 2, seminal ribonuclease, ephrin-A1 and caltrin. Experiment 1 identified and correlated buffalo seminal plasma proteins with sperm motile semen, these proteins being possible markers of sperm quality in the species. Experiment 2 compared the protein profile of the seminal plasma of buffaloes and cattle and found protein differences between species.

Biblioteca responsável: BR68.1