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| Author(s): |
Michelle Silva Araujo
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Botucatu. 2019-08-22. |
| Institution: | Universidade Estadual Paulista (Unesp). Faculdade de Medicina Veterinária e Zootecnia. Botucatu |
| Defense date: | 2018-12-03 |
| Advisor: | Fabiana Ferreira de Souza |
| Abstract | |
Seminal plasma and its proteins influence the sperm quality of chilled and frozen semen in several species, but its influence on the cooled semen of dogs has not yet been clarified. Furthermore, interracial differences in the expression of seminal plasma proteins that may interfere with the quality of canine semen have not been reported. The objective of this study was to verify if the racial pattern interferes in the protein composition of seminal plasma and sperm proteins and to correlate them with the cooling of canine semen in the presence and absence of seminal plasma. In this way, two experiments were performed. In the first experiment the seminal plasma proteins of 5 canine breeds and sperm proteins of 4 breeds were characterized. The semen was collected, evaluated macro and microscopically. Seminal plasma and sperm cells were separated by centrifugation, refrigerated at 5° C and transported to the laboratory for proteomic analysis. Seminal plasma and spermatozoa proteins were digested, submitted to mass spectrometry, identified and characterized for gene ontology. The relationship between the proteins found and the canine breeds was evaluated by multivariate analysis. In the seminal plasma, 10 relevant proteins were found and in the spermatozoa five proteins were considered relevant. Differences were observed in the expression of sperm proteins from different canine breeds. For the second experiment, 3 ejaculate pools of 8 dogs from experiment 1 were used. The morphology, computer-assisted sperm motility (CASA), intact and damaged spermatozoa (eosin / nigrosin), plasma and acrosome membrane integrity, superoxide production, hydrogen peroxide production, mitochondrial potential, lipid peroxidation, oxygen reactive species production (induced and spontaneous thiobarbituric acid technique) and antioxidant enzyme production (ELISA) were evaluated every 48 and 72 hours (M0, M48, M96 and M168) until 168 hours (7 days) in samples of canine semen cooled at 5º C with and without seminal plasma. Seminal samples were divided into two groups, semen extended in TRIS-egg yolk-glucose with seminal plasma and semen extended in TRIS-egg yolk-glucose without seminal plasma. Data were analyzed by two-way ANOVA and multivariate analysis using the Bonferroni test. Seminal plasma refrigerated samples showed high total motility, progressive motility, mean velocity (VAP), straight velocity, rapid spermatozoa, movement index, plasma and acrosomal sperm membranes, greater mitochondrial potential, low lateral head movement and low medium and slow spermatozoa. According to refrigeration time, statistical was significant for total motility, straight velocity, lateral head movement, straightness coefficient, linearity, oscillation coefficient and movement index. The profile of seminal plasma and spermatozoa proteins from the ejaculate of four canine breeds were described. Similarities between Golden Retriever and Maremmano-Abruzzese Sheepdog; and between Great Dane and Bernese Mountain Dog in proteomic profile of spermatozoa exist; while MaremmanoAbruzzese Sheepdog, Gread Dane and Bernese were correlated in proteomic profile of seminal plasma. Seminal plasma exerted a positive influence on the parameters of motility and sperm viability and mitochondrial function during semen refrigeration at 5 ºC. (AU) | |
| FAPESP's process: | 17/02530-8 - Influence of ejaculate proteins from different canine breeds on quality and survival refrigerated sperm with or without seminal plasma |
| Grantee: | Michelle Silva Araujo Volpato |
| Support Opportunities: | Scholarships in Brazil - Doctorate |