KIF3A and KIF3B are two N-terminal motor proteins belonging to the kinesin-II superfamily that play essential roles in spermiogenesis.To understand the roles played by KIF3 A/3B during spermatogenesis of large yellow ...KIF3A and KIF3B are two N-terminal motor proteins belonging to the kinesin-II superfamily that play essential roles in spermiogenesis.To understand the roles played by KIF3 A/3B during spermatogenesis of large yellow croaker Larimichthys crocea,we studied the testis characteristics at different developmental stages of L.crocea,and determined the spatiotemporal expression patterns of kif3a and kif3b during spermiogenesis.Quantitative real-time PCR(qR T-PCR)showed that the overall trends of kif3 a/3 b m RNA abundance during testis development are similar.From stage Ⅱ to stage V,kif3a/3b m RNA abundances first increased and then fell after reaching a peak at stage IV.Interestingly,the m RNA abundances of both genes at stage V were higher than those at stages Ⅱ and Ⅲ.In addition,it is worth of noting that kif3 b m RNA abundance was higher than that of kif3a at all stages.Fluorescence in situ hybridization results revealed that kif3a/3b m RNA abundance dynamics were consistent with the migration of mitochondria,the deformation of nucleus,and the formation of tail.The m RNA hybridization signals of both genes first appeared either around the nuclear periphery or on the side of the nuclei,then appeared at one side of nuclei,and finally were mainly on the tail during spermiogenesis.Our findings contributed to better understanding the molecular mechanisms of spermiogenesis in fish;and suggested that KIF3A and KIF3B may participate in the intracellular transport of mitochondria,nuclear deformation,and the formation of tail during the spermiogenesis in L.crocea.展开更多
Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during sp...Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still un-known.In this study,the cytological features of spermatogenesis were investigated in Larimichthys crocea.In addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also investigated.The full-length cDNA and protein(Lc-PHB)from the L.crocea phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,respec-tively.Lc-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil domains.It was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and brain.Lc-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ hybridization.An immunofluorescence assay revealed that PHB protein was localized in the mitochondria during spermatogenesis.Specifically,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm midpiece.Western blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of testes.Conclusively,these results indicated that PHB was expressed in the mitochondria dur-ing spermatogenesis.In addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.展开更多
基金financially supported by the Scientific and Technical Project of Zhejiang Province (Nos. 2016C02055-7, LY18C190007)the Ningbo Natural Science Foundation (No. 2016A610081)+3 种基金the Scientific and Technical Project of Ningbo (No. 2015C110005)the National Natural Science Foundation of China (No. 31602140)the Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculturethe K. C. Wong Magna Fund in Ningbo University
文摘KIF3A and KIF3B are two N-terminal motor proteins belonging to the kinesin-II superfamily that play essential roles in spermiogenesis.To understand the roles played by KIF3 A/3B during spermatogenesis of large yellow croaker Larimichthys crocea,we studied the testis characteristics at different developmental stages of L.crocea,and determined the spatiotemporal expression patterns of kif3a and kif3b during spermiogenesis.Quantitative real-time PCR(qR T-PCR)showed that the overall trends of kif3 a/3 b m RNA abundance during testis development are similar.From stage Ⅱ to stage V,kif3a/3b m RNA abundances first increased and then fell after reaching a peak at stage IV.Interestingly,the m RNA abundances of both genes at stage V were higher than those at stages Ⅱ and Ⅲ.In addition,it is worth of noting that kif3 b m RNA abundance was higher than that of kif3a at all stages.Fluorescence in situ hybridization results revealed that kif3a/3b m RNA abundance dynamics were consistent with the migration of mitochondria,the deformation of nucleus,and the formation of tail.The m RNA hybridization signals of both genes first appeared either around the nuclear periphery or on the side of the nuclei,then appeared at one side of nuclei,and finally were mainly on the tail during spermiogenesis.Our findings contributed to better understanding the molecular mechanisms of spermiogenesis in fish;and suggested that KIF3A and KIF3B may participate in the intracellular transport of mitochondria,nuclear deformation,and the formation of tail during the spermiogenesis in L.crocea.
基金funded by the Natural Science Foun dation of Zhejiang Province(No.LY18C190007)the Na tural Science Foundation of China-Zhejiang Joint Fund for the Integration of Industrialization and Informatization(No.U1809212)+3 种基金the Natural Science Foundation of Ningbo City(No.2018A610228)the Scientific and Technical Project of Zhejiang Province(No.2021C02069-1)the Scientific and Technical Project of Ningbo City(No.2021Z002),the Scientific Research Foundation of Ningbo University(No.XYL19023)the Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture,the K.C.Wong Magna Fund in Ningbo University.
文摘Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still un-known.In this study,the cytological features of spermatogenesis were investigated in Larimichthys crocea.In addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also investigated.The full-length cDNA and protein(Lc-PHB)from the L.crocea phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,respec-tively.Lc-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil domains.It was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and brain.Lc-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ hybridization.An immunofluorescence assay revealed that PHB protein was localized in the mitochondria during spermatogenesis.Specifically,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm midpiece.Western blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of testes.Conclusively,these results indicated that PHB was expressed in the mitochondria dur-ing spermatogenesis.In addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.
