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热敏离子通道TRPV1在一种淡水硬骨鱼(印度野鲮)的精子中内源性表达,并调节精子活力。

Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility.

作者信息

Majhi Rakesh Kumar, Kumar Ashutosh, Yadav Manoj, Swain Nirlipta, Kumari Shikha, Saha Ashish, Pradhan Avinash, Goswami Luna, Saha Somdatta, Samanta Luna, Maity Apratim, Nayak Tapas Kumar, Chattopadhyay Subhasis, Rajakuberan Chitra, Kumar Abhishek, Goswami Chandan

机构信息

National Institute of Science Education and Research; Institute of Physics Campus; Bhubaneswar, India.

Central Institute of Freshwater Aquaculture; Bhubaneswar, India.

出版信息

Channels (Austin). 2013 Nov-Dec;7(6):483-92. doi: 10.4161/chan.25793. Epub 2013 Aug 2.

Abstract

Sperm cells exhibit extremely high sensitivity in response to slight changes in temperature, osmotic pressure and/or presence of various chemical stimuli. In most cases throughout the evolution, these physico-chemical stimuli trigger Ca (2+)-signaling and subsequently alter structure, cellular function, motility and survival of the sperm cells. Few reports have recently demonstrated the presence of Transient Receptor Potential (TRP) channels in the sperm cells from higher eukaryotes, mainly from higher mammals. In this work, we have explored if the sperm cells from lower vertebrates can also have thermo-sensitive TRP channels. In this paper, we demonstrate the endogenous presence of one specific thermo-sensitive ion channel, namely Transient Receptor Potential Vanilloid family member sub type 1 (TRPV1) in the sperm cells collected from fresh water teleost fish, Labeo rohita. By using western blot analysis, fluorescence assisted cell sorting (FACS) and confocal microscopy; we confirm the presence of this non-selective cation channel. Activation of TRPV1 by an endogenous activator NADA significantly increases the quality as well as the duration of fish sperm movement. The sperm cell specific expression of TRPV1 matches well with our in silico sequence analysis. The results demonstrate that TRPV1 gene is conserved in various fishes, ranging from 1-3 in copy number, and it originated by fish-specific duplication events within the last 320 million years (MY). To the best of our knowledge, this is the first report demonstrating the presence of any thermo-sensitive TRP channels in the sperm cells of early vertebrates as well as of aquatic animals, which undergo external fertilization in fresh water. This observation may have implications in the aquaculture, breeding of several fresh water and marine fish species and cryopreservation of fish sperms.

摘要

精子细胞对温度、渗透压和/或各种化学刺激的轻微变化表现出极高的敏感性。在进化过程中的大多数情况下,这些物理化学刺激会触发钙(2+)信号传导,随后改变精子细胞的结构、细胞功能、运动能力和存活率。最近很少有报告表明高等真核生物,主要是高等哺乳动物的精子细胞中存在瞬时受体电位(TRP)通道。在这项工作中,我们探讨了低等脊椎动物的精子细胞是否也能具有热敏TRP通道。在本文中,我们证明了从淡水硬骨鱼印度野鲮收集的精子细胞中内源性存在一种特定的热敏离子通道,即瞬时受体电位香草酸受体1型(TRPV1)。通过蛋白质免疫印迹分析、荧光辅助细胞分选(FACS)和共聚焦显微镜;我们证实了这种非选择性阳离子通道的存在。内源性激活剂NADA对TRPV1的激活显著提高了鱼精子运动的质量和持续时间。TRPV1在精子细胞中的特异性表达与我们的电子序列分析非常吻合。结果表明,TRPV1基因在各种鱼类中是保守的,拷贝数从1到3不等,它是在过去3.2亿年(MY)内由鱼类特有的复制事件产生的。据我们所知,这是第一份证明早期脊椎动物以及在淡水中进行体外受精的水生动物的精子细胞中存在任何热敏TRP通道的报告。这一观察结果可能对水产养殖、几种淡水和海水鱼类的育种以及鱼精子的冷冻保存具有重要意义。

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