Es-KIF3A Is Involved In The Regulation Of Spermatogenesis Of Eriocheir Sinensis Via Positive Feedback Regulation Of The Wnt/?-catenin Signaling Pathway

The Wnt/?-catenin pathway participates in many important physiological events such as cell proliferation and differentiation in the male reproductive system.We found that Kinesin-2 motor KIF3 A is highly expressed during spermatogenesis in Eriocheir sinensis(E.sinensis),it may potentially promote the intracellular transport of cargoes in this process.However,only a few studies focused on the relationship between KIF3 A and the Wnt/?-catenin pathway in the male reproductive system of decapod crustaceans.In this study,we cloned and characterized the CDS of ?-catenin in E.sinensis for the first time.Fluorescence in-situ hybridization and immunofluorescence results showed the co-localization of Es-KIF3 A and Es-?-catenin at the mRNA and the protein level respectively.To furtherly explore the regulatory function of Es-KIF3 A to the Wnt/?-catenin pathway,the es-kif3 a was knocked down by double strand RNA(dsRNA)in vivo and in primary cultured cells of testes of E.sinensis.Results showed that the expression of es-?-catenin and es-dvl were decreased in the es-kif3 a knockdown group.The protein expression level of Es-?-catenin was also reduced and the location of Es-?-catenin was changed from nucleus to cytoplasm in late stage of spermatogenesis when es-kif3 a was knocked down.Besides,the co-IP result demonstrated that Es-KIF3 A could bind with Es-?-catenin.In summary,this study indicated that Es-KIF3 A can positively regulate the Wnt/?-catenin pathway during spermatogenesis and Es-KIF3 A can bind with Es-?-catenin to facilitate the nuclear translocation of Es-?-catenin.