Avaliação in vitro do efeito dos espermatozoides sobre a expressão gênica em agregados de células epiteliais da tuba uterina bovina

Abstract

The interaction between spermatozoa and uterine tubal cells ensures the availability of viable spermatozoids with a higher fertilization potential. Studies related to sperm binding to the uterine tube have shown that this influence influences the transcription and translation of genes, promoting the formation of an environment conducive to the maintenance of sperm viability. The objective of this study was to evaluate the effect of sperm concentration and incubation time on the relative expression of the FUT 6, NQ01, CST6, B3GNT3, CKB, RARRES2, MIF and FOS genes. For each collection of samples, a sperm pool of six bulls was incubated with BOEC aggregates at concentrations of 1x103 and 1x106 spermatozoids / mL for 18h in a cell culture oven, in addition to the control groups, with only cell aggregates collected at time 0h and 6:00 p.m. Subsequently based on the result of this experiment, it was evaluated whether the co-incubation for 3, 6 and 12h presented variation in the level of BOEC transcripts. Data were analyzed by analysis of variance (ANOVA) and the differences between means were compared by the Tukey test (p <0.05). In relation to the sperm concentration, there was no effect on the relative level of transcripts of genes RARRES2, MIF, CKB, CST6, B3GNT3 and NQO1 in relation to the 0h control without incubation. For the FOS gene there was an increase of its expression by the treatment of 1x106 spermatozoa / mL in 18h of co-incubation with the BOEC. In relation to the incubation time, only the MIF gene presented lower expression after 12 h of coincubation in relation to the moment 0 and 6 h. Based on these results, we conclude that the interaction of spermatozoa and BOEC modulates the expression of the MIF and FOS gene in vitro. For the other evaluated genes, no effect of spermatozoa on the level of transcripts was identified.