睾丸间质细胞特异性表达Cre重组酶转基因小鼠的建立

目的苗勒氏管激素受体-2（anti—mullerianhormonereceptor-2，Amhr2）是睾丸间质细胞中的特异性标志分子，我们构建了Amhr2基因启动子介导的睾丸间质细胞特异性表达Cre重组酶转基因小鼠（Amhr2一Cre）。方法采用显微注射法将7．1kb的转基因片段导入小鼠基因组，获得子代小鼠睾丸组织进行PCR检测，再将转基因小鼠与ROSA26报告小鼠交配，利用LaeZ染色对Amhr2-Cre双转基因进行检测。结果获得子代Amhr2-Cre转基因小鼠经PCR检测显示，睾丸间质细胞中Cre重组酶介导的Amhr2基因发生重组；LacZ染色进一步表明，Cre重组酶在睾丸间质细胞中特异性表达，并介导ROSA位点LoxP序列间的重组。结论建立的睾丸间质细胞特异性表达Cre重组酶的转基因小鼠在睾丸组织中具有一定的组织特异性，并能在体内成功介导这些睾丸间质细胞基因组上LoxP位点间的重组，是一种研制在睾丸特定细胞中特异性基因剔除小鼠的良好遗传工具小鼠。

Establishment of testicle leydig cells specific Cre transgenic mice.

Objetive Amhr 2 protein is the specific protein of testicle tissue. We generated a new transgenic line expressing Cre recombinase under the control of an Amhr 2 gene promoter （ Amhr 2-Cre） to in- vestigate sexual development. Methods Microinjeetion was employed to introduce the 7. I kb transgenic frag- ment into oocytes,and progenies were obtained. PCR detected expression of Amhr 2-Cre transgene within testi- cle tissue containing leydig cells. Transgenie line was crossed with ROSA26 report line, the activity of Cre reeombinase was revealed by LacZ staining in ROSA26; Amhr 2-Cre double transgenic mice. Results PCR products revealed that tile Amhr 2 gene was deleted by Cre mediated recombination in the leydig cells. LacZ staining of tile Amhr 2-Cre and ROSA26 double transgenic mice showed that Cre recombinase activity was de- tected in leydig cells. Conclusion All these data indicated that the Cre recombinase has been expressed in the testicle tissues of the Amhr 2-Cre transgenic mice. The mice generated could serve as a useful tool for gener- ating testicle specific gene-knockout mice.