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目的研究新生小鼠睾丸组织异体异位移植后,几种在睾丸支持细胞中起重要作用的基因和蛋白表达情况,为异体异位睾丸组织移植模型用于科研及临床的可行性提供进一步实验数据。方法将162只1～2d昆明小鼠的睾丸移植到54只7～12周去势雄性免疫缺陷小鼠背部;在移植后9个时间段(3d和1～8周)取出移植物;选取4种在睾丸支持细胞中表达或高表达的基因abp、amh、vim和clu,采用聚合酶链反应,对发育不同阶段移植物中4种基因的表达情况进行分析,并与正常小鼠相应各年龄段睾丸中的基因表达相比较;同时采用免疫组织化学方法对支持细胞的GATA-4蛋白在移植物组织中的表达量及分布情况进行分析。结果在9个时间段取出的移植物中,所测定4种基因的表达趋势与在正常小鼠睾丸中所见基本相同;免疫组化结果显不,4周和8周移植物支持细胞中GATA-4蛋白呈高表达,与在正常小鼠睾丸组织支持细胞中的表达基本一致。结论新生小鼠睾丸组织异体异位移植到免疫缺陷小鼠体内后,支持细胞的发育在组织形态学以及几种受试基因的表达趋势和蛋白的表达情况与在正常小鼠中的表现基本相同。
Objective To study the expression of several genes and proteins that play an important role in the testicular sertoli cells after allotransplantation of testicular tissue in newborn mice and provide further experimental data for the feasibility of the allogenic ectopic testicular tissue transplantation model for research and clinical application . Methods Totally 162 testicular Kunming mice were transplanted into the back of 54 castrated male immunodeficient mice for 7-12 weeks. The grafts were removed in 9 time periods (3d and 1-8 weeks) after transplantation. Four The genes abp, amh, vim and clu, which were expressed or highly expressed in testicular sertoli cells, were analyzed by polymerase chain reaction (PCR). The expressions of four genes in different stages of development were analyzed and compared with those of normal mice The expression of GATA-4 protein in the grafted cells was analyzed by immunohistochemical method. Results The expression of the four genes in the grafts removed in 9 time periods was almost the same as that seen in the testis of normal mice. The results of immunohistochemistry showed that the expression of GATA in the grafts after 4 weeks and 8 weeks -4 protein was overexpressed, which was consistent with the expression in normal mouse testis tissue. Conclusions After the allogeneic heterotopic transplantation of testicular tissue of neonatal mice into immunocompromised mice, the development of supporting cells is almost the same in histomorphology and the tendency of expression of several tested genes and protein expression as in normal mice .