The Expression And Function Of HnRNPU In Mouse Testes

Part 1 The expression and localization of hnRNPU in testes?Objective? Study the conservation of hnRNPU between different species and explore the expression level and cellular localization of hnRNPU in both mouse and human testes.? Methods ? Bioinformatics analysis,immunofluorescence,immunohistochemistry,RT-q PCR and Western blot were used in this study.?Results?hnRNPU is a highly conserved protein and the homology of hnRNPU amino acid was ~97.25% between humans and mice.The hnRNPU protein was highly expressed in human and mouse testes where mainly localized in the nuclei of testicular germ cells and Sertoli cells.?Conclusions? hnRNPU is highly conserved proteins in the testes of mammals and localized in the nucleus of testicular germ cell and Sertoli cells.Part 2 The function of hnRNPU in germ cell on spermatogenesis and male fertility in mice?Objective? To study the effects of hnRNPU in germ cells on spermatogenesis and male fertility.? Methods ? The Cre-Loxp system was used to generate the mouse model of conditional knockout Hnrnpu gene in germ cells.The role and function of hnRNPU in spermatogenesis and male fertility were analyzed by fertility test and testicular morphology analyses.?Results? Stra8-Cre mediated conditional knockout of hnRNPU mouse model was successfully generated.The male conditional KO mice were displayed fertility decline and the spermatogenesis in Stra8-Cre-c KO mice was disrupted.?Conclusions? hnRNPU in germ cells plays an important role in spermatogenesis.Ablation of hnRNPU in germ cells leads to abnormal spermatogenesis in mice,and reduce male fertility.Part 3 The function of hnRNPU in Sertoli cells on spermatogenesis and male fertility in mice?Objective? To investigate the role and mechanism of hnRNPU in Sertoli cells on spermatogenesis and Sertoli cell development in mice.?Methods? Cre-Loxp system was used to generated a mouse model of conditional knockout Hnrnpu gene in Sertoli cells.Fertility test,testis morphological analyses,immunofluorescence and immunoprecipitation were used to reveal the role and mechanism of hnRNPU in spermatogenesis and male fertility.? Results ? Amh-Cre mediated hnRNPU conditional knockout mouse model was successfully constructed.Knockout male mice were completely infertile,characterized by impaired testicular development,abnormal Sertoli cell development and proliferation,and spermatogenic cell apoptosis.hnRNPU was co-localized with WT1 in mouse testis and interacted with each other to form complexes.? Conclusions ? hnRNPU in Sertoli cells plays an important role in regulating the development of Sertoli cells and spermatogenesis.The loss of hnRNPU may lead to abnormal Sertoli cell development,abnormal cell structure and morphology,and affect the proliferative ability of Sertoli cell,thus destroying the spermatogenic microenvironment,thereby affecting the formation of seminiferous tubules and spermatogenesis,and ultimately affecting the development of the whole testis and leading to complete male infertility.