Naz R K, Ahmad K, Kaplan P
Reproductive Immunology and Molecular Biology Labs, Albert Einstein College of Medicine, Bronx, New York 10461.
Biol Reprod. 1993 Apr;48(4):720-8. doi: 10.1095/biolreprod48.4.720.
The presence and roles of mitotic cyclins (cyclin A and cyclin B1), and cdc2 and related (having an N-terminal PSTAIRE conserved sequence) serine/threonine protein kinases were investigated by use of specific antibodies. The cyclin A and cyclin B1 antibodies reacted specifically with the acrosomal regions of human sperm cells in the indirect immunofluorescence technique (IFT) and recognized the specific band of p60 (cyclin A) and p62 (cyclin B1) on the Western blot of sodium deoxycholate (DOC)-solubilized noncapacitated human sperm preparation. Both antibodies reacted more strongly with the specific cell region/band of capacitated sperm than with that of noncapacitated sperm. The cdc2 and PSTAIRE antibodies also reacted predominantly with the acrosomal regions of human sperm cells in IFT and recognized the specific band of 34 kDa corresponding to p34 cdc2 protein on the Western blot of DOC-solubilized noncapacitated human sperm preparation. Again, both antibodies reacted more strongly with the specific cell region/band of capacitated sperm than with that of noncapacitated sperm. The cyclin A antibodies (but not the cyclin B1 antibodies) and cdc2 antibodies as well as the PSTAIRE antibodies significantly (p = 0.02 to p < 0.001) increased (rather than decreased) the human sperm penetration rates of zona-free hamster ova; the cyclin A and cdc2 antibodies showed the strongest enhancing effects. These three antibodies significantly increased the acrosome reaction and release of acrosin activity from the sperm cells.(ABSTRACT TRUNCATED AT 250 WORDS)
利用特异性抗体研究了有丝分裂周期蛋白(周期蛋白A和周期蛋白B1)以及cdc2和相关(具有N端PSTAIRE保守序列)的丝氨酸/苏氨酸蛋白激酶的存在及作用。在间接免疫荧光技术(IFT)中,周期蛋白A和周期蛋白B1抗体与人精子细胞的顶体区域发生特异性反应,并在脱氧胆酸钠(DOC)溶解的未获能人精子制剂的蛋白质免疫印迹中识别出p60(周期蛋白A)和p62(周期蛋白B1)的特异性条带。两种抗体与获能精子的特定细胞区域/条带的反应均强于未获能精子。在IFT中,cdc2和PSTAIRE抗体也主要与人精子细胞的顶体区域发生反应,并在DOC溶解的未获能人精子制剂的蛋白质免疫印迹中识别出对应于p34 cdc2蛋白的34 kDa特异性条带。同样,两种抗体与获能精子的特定细胞区域/条带的反应均强于未获能精子。周期蛋白A抗体(而非周期蛋白B1抗体)、cdc2抗体以及PSTAIRE抗体显著(p = 0.02至p < 0.001)提高(而非降低)了人精子对去透明带仓鼠卵的穿透率;周期蛋白A和cdc2抗体的增强作用最强。这三种抗体显著增加了精子细胞的顶体反应和顶体蛋白酶活性的释放。(摘要截短至250字)