大鼠睾丸发育过程基因表达的研究

本研究选取１、３ 和８（成年）周龄等不同发育阶段大鼠，应用睾丸组织树脂包埋形态学观察与ｍ ＲＮＡ 差异显示逆转录－聚合酶链反应技术（ｄｉｆｆｅｒｅｎｔｉａｌｄｉｓｐｌａｙ ＲＴ－ＰＣＲ，ＤＤＲＴ－ＰＣＲ）相结合，探讨大鼠睾丸发育过程中，精子发生过程睾丸特异基因的表达。结果：对不同周龄大鼠睾丸ｍ ＲＮＡ 进行差异比较，共得到 ８２ 个ｃＤＮＡ 差异片段。又分别标记不同周龄大鼠睾丸全部 ｍ ＲＮＡ 为探针，对其中 ４０ 个ｃＤＮＡ差异片段进行斑点杂交，得到１２ 个初步鉴定的特异或表达增加的ｃＤＮＡ 差异片段，片段范围２５０～５００ ｂｐ，其中２ 个在１ 周龄睾丸组获得，４ 个在３ 周龄睾丸组获得，在８ 周龄组获得 ５ 个，一个为１ 周和３ 周龄共有而８ 周龄大鼠缺乏的ｃＤＮＡ 片段。通过形态学观察与ｍ ＲＮＡ 差异显示的结果对应分析，初步认为２ 个１ 周龄的特异基因片段属于睾丸二倍染色体生精细胞表达基因，４ 个３ 周龄睾丸特异基因片段也属二倍或四倍体生精细胞表达基因，其中Ｂ型精原细胞和初级精母细胞的表达基因占主要成分；而成年动物的５ 个特异基因片段是属单倍体的精子细胞以及减数分裂过程的精母细胞表达基因片段

The study of specific gene expression in the testicular development process of rat

To find the specific gene expression for regulating mitosis and meiosis of germ cells during spermatogenesis of rat testis. Methods: one, three and eight weeks old SD rats were used in the studies. The morphological observation of testicular tissues embedded by resin and mRNA differential display RT PCR(DDRT PCR) were combined to obtain the specific mRNA expression in testicular tissue of 1,3 and 8 weeks old rats. Dot blot hybridization was used to further screen the differential cDNA segments. Results: Eighty two differential cDNA segments were obtained through primary DDRT PCR, among which forty differential cDNA segments were selected for further screening with dot blot hybridization. Twelve primary differential cDNA segments have been screened by the dot blot hybridization. The size of cDNA segments ranged 250 to 500 bp. Two cDNA segments in 1 week old rat, 4 in 3 week old rat and 5 in adult rat testicular tissues were obtained. In addition, one cDNA segment was found in both 1 and 3 week old rat testicular tissue. Conclusion: Two cDNA segments obtained in 1 week old rat were derived from spermatogonia; 4 cDNA segments obtained in 1 week old rat derived either from spermatogonia from primary spermatocytes; 5 cDNA segments obtained in adult rat derived from either spermatids or pachytene spermatocytes in accordance with the morphological analysis as well as the DDRT PCR techniques.