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首页> 外文期刊>Journal of Cellular Physiology >A novel epididymal quiescence factor inhibits sperm motility by modulating NOS activity and intracellular NO‐cGMP pathway
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A novel epididymal quiescence factor inhibits sperm motility by modulating NOS activity and intracellular NO‐cGMP pathway

机译：小说附睾的沉默抑制因素通过调节NOS活性和精子的运动性细胞内没有cGMP通路

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摘要

Mature and potentially motile spermatozoa stored in cauda epididymis in an inactive state for approximately 30 days; however, during ejaculation they regain motility. To understand the actual molecular mechanism of the sperm quiescence during caudal stay, a proteinaceous quiescence factor (QF) has been purified from caprine epididymal plasma to apparent homogeneity. In the present study complete purification, detailed characterization as well as mechanistic pathway of QF has been described. QF is purified to 215‐fold with 45% activity recovery. It is a 59?kDa monomeric protein with isoelectric point 5.8 and optimally active at pH 7.5. Circular dichroism spectroscopy and atomic force microscopy study confirm its α‐helical secondary structure and globular tertiary conformation. QF is a thermo‐stable protein as higher temperature does not alter its helical structure. N‐terminal amino acid sequencing and MALDI analysis of QF did not find 100% similarity with any available protein of the database, proved its novelty. QF at 2?μM dose inhibits sperm progressive forward motility within 10?min. This motility inhibitory activity of QF is mediated by reducing NOS enzyme activity and subsequently decreasing the intracellular NO and cGMP concentration. It does not modulate intracellular Ca ++ and cAMP concentration. QF has no adverse effect on DNA integrity and morphology of spermatozoa. Motility inhibitory action of QF is reversible. Thus, the role of QF in maintaining energy saving quiescence state of mature cauda spermatozoa and its reactive nitrogen species reducing activity may lead to a new direction for storage of spermatozoa and idiopathic male infertility.
机译：成熟的和潜在的能动的精子存储在尾附睾处于不活跃状态大约30天;他们恢复运动性射精。实际的分子机制的精子蛋白质的静止期间尾静止因子(QF)被净化山羊的附睾的等离子体明显同质性。净化、详细描述《被描述的机械化的途径。QF - 215进行净化褶皱与45%的活动复苏。等电点5.8和最佳活跃在pH值7.5. 力显微镜研究证实其α螺旋二级结构和球状三级构象。更高的温度不会改变它的螺旋结构。QF谱技术分析没有发现100%的相似性与任何可用的蛋白质数据库的,证明其新颖性。精子向前进步的能动性在10 ?分钟。QF这种运动性抑制活动通过减少NOS酶活性和介导的随后减少细胞内没有环鸟苷酸浓度。细胞内钙+ +和营地的浓度。对DNA完整性和没有不良影响呢精子的形态。QF作用是可逆的。在维护节能的静止状态成熟的尾精子及其活性氮可能导致物种减少活动新的存储精子和方向特发性男性不育。

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