Effects of a Sertoli cell specific knockout of the connexin43-gene on the regulation of spermatogenesis in transgenis mice using the Cre/IoxP-recombination system

使用 Cre/IoxP 重组系统，支持细胞特异性敲除 connexin43 基因对转基因小鼠精子发生调节的影响

• 批准号: 45912711
• 负责人: Professor Dr. Ralph Brehm
• 金额: --
• 依托单位: Anatomisches Institut
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/45912711?language=en
• 关键词: Effects; Sertoli; cell; specific; knockout

The predominant testicular gap junctional protein connexin43 (cx43) is located « between neighboring Sertoli cells (SC) and between SC and germ cells. It is involved in • testicular development, germ cell differentiation, initiation and maintenance of spermatogenesis. As total disruption of the cx43 gene leads to perinatal death, we generated a « conditional cx43 knockout (KO) mouse using the Cre/loxP recombination system which | lacks the cx43 gene solely in SC (SCCx43KO) to evaluate the SC specific functions of cx43 on spermatogenesis in-vivo. Adult SCCx43KO~A mice showed normal testis descent and • development of the urogenital tract, but testis size and weight was drastically lower when | compared with heterozygous and wild-type (WT) littermates. Histological analysis and quantitation of mRNA expression of germ cell-specific marker genes revealed a significant , • reduction in the number of spermatogonia with only a few tubules left showing normal spermatogenesis. Thus, SC-specific deletion of cx43 mostly .resulted in an arrest of spermatogenesis at the level of spermatogonia or SC-only syndrome and to intratubular SC • clusters. Our data demonstrate that cx43 expression in SC is an absolute requirement for normal testicular development and spermatogenesis.

主要的睾丸间隙连接蛋白连接蛋白43（cx43）位于相邻的支持细胞（SC）之间以及SC和生殖细胞之间。它参与睾丸发育、生殖细胞分化、精子发生的启动和维持。由于cx43基因的完全破坏导致围产期死亡，我们使用Cre/loxP重组系统产生了条件性cx43敲除（KO）小鼠，|在SC中单独缺失cx43基因（SCCx 43 KO），以评估cx43在体内精子发生中的SC特异性功能。成年SCCx 43 KO ~A小鼠表现出正常的睾丸下降和泌尿生殖道发育，但睾丸大小和重量显著降低，|与杂合子和野生型（WT）同窝仔相比。组织学分析和生殖细胞特异性标记基因的mRNA表达的定量揭示了精原细胞数量的显著减少，仅剩下少数小管，显示正常的精子发生。因此，SC特异性的cx43缺失主要导致精原细胞水平的精子发生停滞或仅SC综合征和小管内SC簇。我们的数据表明，SC中的cx43表达是正常睾丸发育和精子发生的绝对要求。