The Localization And Expression Of Phosphotyrosine Proteins On Cervus Elaphus Spermatozoa In Vitro Capacitation

Mammalian spermatozoa capacitation is a series of complicated events. They must undergo a post-ejaculatory period of maturation, known as capacitation, before they can engage in the process of fertilization. Many factors were involved in the regulation of the capacitation, and tyrosine phosphorylation plays an important role in the regulation of the the movement, hyperactivated motility and the capacitation. The molecular basis of mammalian sperm capcitation is poorly understood. It is well known that sperm capacitation is associated with an increase in tyrosine phosphorylation of a subset of proteins. The aim of this study was to evaluate the change of tyrosine phosphorylation on the red deer spermatozoa in vitro during capacitation.The spermatozoa were incubated in modified TALP under 5% CO2 in air at 37℃.The capacitation effect was assessed by chlortetracycline (CTC) staining. Indirect immunofluorescence and western blotting were used to analyze the level and localization of tyrosine phosphorylation. The results show that the protein tyrosine phosphorylation increase during the in vitro capacitation. The distribution of the phosphotyrosine-specific fluorescence amplify from the head of sperm to the sperm flagellum. And western blot analysis shows that there are 6 kinds of phosphotyrosine proteins after capacitation, while there only 1 phosphotyrosine proteins before incubation.So we can say that protein tyrosine phosphorylation inereased during in vitro capacitation and there may be a possible correlation between protein tyrosine phosphorylation.