摘要
兔附睾尾液经ＨＰＬＣ纯化后，获得分子量为２０ｋＤ的附睾特异蛋白，命名为ＢＥ－２０。ＢＥ－２０的抗体具有明显抑制人精子穿透去透明带仓鼠卵的能力。已获得编码ＢＥ－２０的０．５ｋｂｃＤＮＡ片段，命名为ＲＥＤ－２０。用地高辛标记此片段，与兔睾丸、附睾、输精管和肝组织进行原位杂交，观察ＢＥ－２０在各种组织中的定位和表达。采用杂交前热变性与不用热变性组织两种方法进行原位杂交。杂交信号均为紫褐色的颗粒，分布在附睾尾的主细胞和近端输精管的上皮细胞的胞质内，表明胞质内ｍＲＮＡ与地高辛标记ＲＥＤ－２０ｃＤＮＡ探针杂交。组织经热变性后，附睾尾和近端输精管上皮细胞核出现强阳性反应，表明核内的同源ＤＮＡ与地高辛标记的ＲＥＤ－２０ｃＤＮＡ探针杂交。但在睾丸和肝脏内未见杂交信号，提示ＢＥ－２０是附睾尾和近端输精管上皮细胞合成的蛋白质。经分析证明，此蛋白是一种细胞外的蛋白酶抑制剂，可能与防止精子过早获能有关，它是否能用于制备抗生育疫苗有待深入的研究。
An epididymal specific protein with molecular weight 20 kD of rabbit was purified by HPLC technique and named BE-20 protein. The antibody against to BE 20 protein was able to inhibit the capacity of human sperm penetration into zona pellucida-free hamster egg. The BE-20 protein is a kind of glycoproteins, The 0.5 kb cDNA coding for BE-20 protein was obtained and labeled with digoxigenin DNA labeling kit.In situ hybridization of testis, epididymis,vas deferens and liver tissues of rabbit were performed in order to localize BE20 mRNA and DNA distribution. Both the denatured and non-denatured tissues were approached.The purple brown granules of hybridized signal distributed in the cytoplasm of principle cells of cauda epididymides. After denature of tissues,the potential positive reaction granules were found to be distributed in the nuclei of epithelial cells of proximal vas deferens due to homogenous nucleic DNA with dig-DNA probe hybridization. But there are no hybridization signals in testis and liver tissues. The present study demonstrated that the specific epididymis protein of rabbit BE-20 is expressed in the epithelia of both the cauda epididymides as well as the vas deferens of rabbit.
出处
《解剖学报》
CAS
CSCD
北大核心
1995年第1期53-57,T010,11,共7页
Acta Anatomica Sinica
基金
国家86.3高科技项目
关键词
附睾蛋白BE-20
基因表达
原位杂交
Epididymis specific protein BE-20
Gene expression
In situ hybridization
Rabbit
