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人精子体外获能过程中线粒体呼吸效率的评估

Evaluation of mitochondrial respiratory efficiency during in vitro capacitation of human spermatozoa.

作者信息

Stendardi A, Focarelli R, Piomboni P, Palumberi D, Serafini F, Ferramosca A, Zara V

机构信息

Applied Biology Section, Department of Biomedical Sciences, University of Siena, Siena, Italy.

出版信息

Int J Androl. 2011 Jun;34(3):247-55. doi: 10.1111/j.1365-2605.2010.01078.x.

Abstract

The role of mitochondria in sperm motility was the subject of several investigations. However, different views on this topic emerged among scientists. In particular, very little is known on the mechanisms of energy production occurring during human sperm capacitation and related processes. In this study, we have investigated the mitochondrial respiratory efficiency in human sperm samples from normozoospermic subjects before and after swim-up selection and incubation under capacitating condition. Sperm cells, selected by swim-up treatment, were incubated up to 24 h and then demembranated by hypotonic swelling at selected times. The oxygen uptake rate was measured in both basal and swim-up selected samples by a polarographic assay. Mitochondria of swim-up selected cells showed an impressive oxygen consumption rate, which was about 20 times higher than that measured in basal samples. The high mitochondrial respiratory efficiency remained stable up to 24 h after the swim-up treatment. The respiration control ratio, the substrate specificity and the inhibitor sensitivity in the swim-up selected samples were similar to those of basal samples thereby suggesting that the physiology of mitochondria was preserved after the swim-up treatment. Furthermore, the remarkably high mitochondrial respiration in swim-up selected samples allowed the oxygraphic analysis of just 200,000 sperm cells. Sperm selection and incubation under capacitating condition are therefore associated with a high activity of the mitochondrial respiratory chain. The sperm oxygen consumption rate could be useful to exclude mitochondria malfunctioning in male infertility.

摘要

线粒体在精子运动中的作用是多项研究的主题。然而,科学家们对这一话题出现了不同的观点。特别是,关于人类精子获能及相关过程中能量产生的机制,人们了解甚少。在本研究中,我们调查了正常精子受试者的人类精子样本在游动选择前后以及在获能条件下孵育后的线粒体呼吸效率。通过游动处理选择的精子细胞孵育长达24小时,然后在选定时间通过低渗肿胀使其去膜。通过极谱分析测量基础样本和游动选择样本中的氧摄取率。游动选择细胞的线粒体显示出令人印象深刻的氧消耗率,比基础样本中测得的氧消耗率高约20倍。在游动处理后长达24小时,高线粒体呼吸效率保持稳定。游动选择样本中的呼吸控制率、底物特异性和抑制剂敏感性与基础样本相似,这表明游动处理后线粒体的生理功能得以保留。此外,游动选择样本中极高的线粒体呼吸使得仅对20万个精子细胞进行氧电极分析成为可能。因此,在获能条件下进行精子选择和孵育与线粒体呼吸链的高活性相关。精子氧消耗率可能有助于排除男性不育中线粒体功能障碍。

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