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CATSPER1在人睾丸及射出精子中的表达及意义

The expression and significance of CATSPER1 in human testis and ejaculated spermatozoa.

作者信息

Li Hong-Gang, Liao Ai-Hua, Ding Xiao-Fang, Zhou Hui, Xiong Cheng-Liang

机构信息

Centre of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Asian J Androl. 2006 May;8(3):301-6. doi: 10.1111/j.1745-7262.2006.00132.x.

Abstract

AIM

To investigate the distribution of cation channel of sperm 1 (CATSPER1) protein and the presence of CATSPER1 mRNA in human testis and ejaculated spermatozoa. The influence of anti-human CATSPER1 antibody upon human sperm motility was used to evaluate the function of human CATSPER1 and to estimate its possible use as a target for immunocontraception.

METHODS

Human ejaculated sperm from normozoospermic donors (n = 12) and liquid nitrogen frozen human testis were used for the study of mRNA and protein expression of CATSPER1 by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively. Spermatozoa from normozoospermic donors (n = 12) were individually processed using a swim-up procedure and were then incubated with CATSPER1 antibody at final concentrations of 20, 4 and 0.8 microg/mL. After 1, 2 and 6 h incubation, progressive motility and fast progressive motility were measured by means of computer-assisted semen analysis.

RESULTS

CATSPER1 transcript was detected in both human testis and each human ejaculated semen sample. CATSPER1 protein expressed in the membrane of spermatid and was localized in the principal piece of the sperm tail. The application of CATSPER1 antibody at all concentrations significantly inhibited both progressive motility and fast progressive motility after 1, 2 and 6 h incubation, and significant dose-dependent changes were observed.

CONCLUSION

CATSPER1 is meiotically and post-meiotically expressed in human testis tissue. CATSPER1 mRNA in human ejaculated spermatozoa could be a more feasible target for study and infertility screening than testis biopsy. In addition, our results suggest that human CATSPER1 could be a possible target for immunocontraception.

摘要

目的

研究精子阳离子通道1(CATSPER1)蛋白在人睾丸和射出精子中的分布以及CATSPER1 mRNA的存在情况。利用抗人CATSPER1抗体对人精子活力的影响来评估人CATSPER1的功能,并估计其作为免疫避孕靶点的潜在用途。

方法

分别采用逆转录聚合酶链反应(RT-PCR)和免疫组织化学方法,对来自正常精子捐献者(n = 12)的人射出精子以及液氮冷冻保存的人睾丸进行研究,以检测CATSPER1的mRNA和蛋白表达。对来自正常精子捐献者(n = 12)的精子采用上游法单独处理,然后与终浓度为20、4和0.8μg/mL的CATSPER1抗体孵育。孵育1、2和6小时后,通过计算机辅助精液分析测量精子的前向运动能力和快速前向运动能力。

结果

在人睾丸和每份人射出精液样本中均检测到CATSPER1转录本。CATSPER1蛋白在精子细胞的细胞膜中表达,且定位于精子尾部的主段。在所有浓度下,CATSPER1抗体孵育1、2和6小时后均显著抑制了精子的前向运动能力和快速前向运动能力,且观察到显著的剂量依赖性变化。

结论

CATSPER1在人睾丸组织减数分裂期和减数分裂后期均有表达。与睾丸活检相比,人射出精子中的CATSPER1 mRNA可能是更可行的研究和不育筛查靶点。此外,我们的结果表明人CATSPER1可能是免疫避孕的一个潜在靶点。

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