【摘要】 目的:比较成年大鼠睾丸生精小管处于减数分裂期的发育片段和睾丸间质细胞所表达基因的差异,初步筛选出与减数分裂相关的基因,为进一步研究减数分裂相关基因对精子发生的调控奠定基础。方法:运用在透射光解剖显微镜下区分和显微分割的方法,将成年大鼠新鲜睾丸生精小管第X III~I期处于减数分裂阶段的片段分离出来,同时分离睾丸间质细胞,将二者进行mRNA差异显示逆转录聚合酶链反应(DDRT-PCR)分析,所得差异片段进行纯化回收,然后进行反向斑点杂交。结果:经mRNA差异显示,第X III~I期生精小管片段共回收到7个差异cDNA片段,而间质细胞共回收到9个差异cDNA片段。经反向斑点杂交,共获得11个初步鉴定的特异表达增加的差异cDNA片段,片段大小为200~500 bp,其中6个从第X III~I期生精小管片段获得,5个从间质细胞获得。结论:这些差异片段可作为睾丸减数分裂表达的序列标签进行更深入地研究。更多还原

【Abstract】 Objective: To find the expression of specific genes related to the meiosis of germ cells during spermatogenesis in the rat testis.Methods: Segments of seminiferous tubules were obtained from the adult male SD rats,at stages XIII~I of meiosis,and the interstitial cells of the same testis were isolated under the stereomicroscope.The total RNAs of stages XIII～I segments and the testicular interstitial cells were extracted respectively,and mRNA differential display RT-PCR(DDRT-PCR) was conducted.The obtained cDNA fractions were purified and recovered,the reverse dot blot hybridization,sub-clones and screens of blue/white dots performed,and the results of sub-clones were identified by restriction endonuclease EcoR I digestion.Results: Sixteen differential cDNA fractions were obtained through primary DDRT-PCR,7 from stages XIII～I segments and 9 from the testicular interstitial cells.Another 11 were selected for further screening by reverse dot blot hybridization,their size ranging from 200 to 500 bp,of which 6 were from the stages(XIII)～I segments of seminiferous tubules and the other 5 from the rat testicular interstitial cells.All of the 11 cDNA fractions were sub-cloned and screened by blue/white dots.Conclusion: Specifically expressed differential cDNA fractions can be obtained and primarily identified from testicular interstitial cells and the seminiferous tubules,which,as the sequence tags of the testicular meiotic expression,deserve further investigation.更多还原