Li Shifeng, Zhang Yunbin, Liu Haixiong, Yan Yuanchang, Li Yiping
Shanghai Key Laboratory for Molecular Andrology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China.
Acta Biochim Biophys Sin (Shanghai). 2008 Aug;40(8):761-7.
Our previous studies showed that gamma-aminobutyric acid (GABA)A and GABAB receptors are involved in rat sperm acrosome reaction induced by progesterone or GABA. Here, we report the presence of GABAC receptor in rat testis and spermatozoa. Full-length complementary DNA encoding the rho1, rho2 and rho3 subunits of GABAC receptor were cloned from rat testis; their sequences are identical to those of rat GABAC receptor in retina. Reverse transcription-polymerase chain reaction analysis showed that during the development of rat testis, the transcript levels of the rho1 and rho2 subunits showed little change, while the expression of rho3 was gradually up-regulated. Immunofluorescence analysis using an anti-rho1 antibody revealed that GABAC receptor exists on the elongated spermatid and sperm. Using a chlortetracycline assay, we found that N(4)-chloroacetylcytosine arabinoside, a GABAC receptor agonist, triggered rat sperm acrosome reaction; whereas (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid, a GABAC receptor antagonist, inhibited the ability of N(4)-chloroacetylcytosine arabinoside to induce acrosome reaction. These results suggested that GABAC receptors are also involved in rat sperm acrosome reaction.
我们之前的研究表明,γ-氨基丁酸(GABA)A和GABAB受体参与了孕酮或GABA诱导的大鼠精子顶体反应。在此,我们报告大鼠睾丸和精子中存在GABAC受体。从大鼠睾丸中克隆出编码GABAC受体rho1、rho2和rho3亚基的全长互补DNA;它们的序列与大鼠视网膜中的GABAC受体序列相同。逆转录-聚合酶链反应分析表明,在大鼠睾丸发育过程中,rho1和rho2亚基的转录水平变化不大,而rho3的表达则逐渐上调。使用抗rho1抗体的免疫荧光分析显示,GABAC受体存在于延长型精子细胞和精子上。使用金霉素检测法,我们发现GABAC受体激动剂N(4)-氯乙酰胞嘧啶阿拉伯糖苷引发了大鼠精子顶体反应;而GABAC受体拮抗剂(1,2,5,6-四氢吡啶-4-基)甲基次膦酸抑制了N(4)-氯乙酰胞嘧啶阿拉伯糖苷诱导顶体反应的能力。这些结果表明,GABAC受体也参与了大鼠精子顶体反应。