Patterns of tyrosine phosphorylation in boar epididymal and ejaculated spermatozoa capacitated in vitro | Patrones de fosforilación de tirosina en espermatozoides porcinos, epididimarios y eyaculados, capacitados in vitro
2011
Avilés López, K., Universidad de Murcia (España). Facultad de Veterinaria | García Vázquez, F.A., Universidad de Murcia (España). Facultad de Veterinaria | Gadea, J., Universidad de Murcia (España). Facultad de Veterinaria | Matas, C., Universidad de Murcia (España). Facultad de Veterinaria
The aim of this study was to determine the P-tyrosine phosphorylation pattern in epididymal (EP) and ejaculated (EJ) spermatozoa before and after capacitation. Sperm samples were processed by Percoll gradient selection and incubation in TALP media up to 3 hours. After immunofluorescence staining we evaluated that presence of tyrosine phosphorilation was characterized in boar spermatozoa by a fluorescent signal presents in the equatorial subsegment, triangular in appearance. Before capacitation, the EJ spermatozoa presented a higher percentage of no phosphorylation than EP cells (EJ 84.13+/-5.53% vs. EP 32.00+/-7.35%, P<0.01). After capacitation, the proportion of phosphorylated sperm increased in EP groups and this high level of phosphorylation was maintained from 0 to 3 hours of incubation. For EJ group at 3 h of co incubation has an increased of phosphorylated proteins in the group III and a decrease in the group II (42,13+/-16,66). The differences between EP and EJ before capacitation suggest that some components of the seminal plasma are implied in the sperm membrane stabilisation. However, the capacitation process is equally effective for both EP and EJ spermatozoa to induce phosphorylation signals. These results confirmed the previously results obtained by Western-blot analysis of the same samples.
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