Boar sperm tyrosine phosphorylation patterns in the presence of oviductal epithelial cells: in vitro, ex vivo, and in vivo models

Spermatozoa transport through the oviduct is a controlled process that regulates sperm capacitation. A crucial event involved in capacitation is protein tyrosine phosphorylation (TP). This study was undertaken to determine whether similarities exist in protein TP distribution between spermatozoa bound or unbound to oviductal epithelial cells (OEC) in three different conditions; i) in vitro, spermatozoa coincubated with OEC cultures; ii) ex vivo, spermatozoa deposited in porcine oviductal explants from slaughtered animals; iii) in vivo, in which sows were inseminated and the oviduct was recovered. The localization of phosphotyrosine protein was determined using indirect immunofluorescence. The distribution of protein TP was significantly (P<0.05) different between bound and unbound cell populations in all experiments. In sows inseminated close to ovulation, spermatozoa were found mainly in the utero tubal junction, where spermatozoa exhibited higher proportion of flagellum phosphorylation. Spermatozoa not bound to OEC exhibited high levels of protein phosphorylation (phosphorylated equatorial subsegment and acrosome and/or phosphorylated flagellum) in the ex vivo and in vivo experiments (P<0.05). However, unbound spermatozoa coincubated with OEC in in vitro conditions tended to show intermediate levels of TP (equatorial subsegment with or without phosphorylated flagellum). In spermatozoa bound to OEC, protein TP was located in the equatorial subsegment or presented no phosphorylation (P<0.05). Although sperm capacitation conditions in vivo were not reproducible in vitro in our experimental conditions, sperm and OEC binding seemed to be a mechanism for selecting spermatozoa with a low level of TP in in vivo, ex vivo, and in vitro experiments.