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检测牛精子中神经生长因子(NGF)及其特异性受体(TrkA),以及 NGF 对精子功能的影响。

Detection of nerve growth factor (NGF) and its specific receptor (TrkA) in ejaculated bovine sperm, and the effects of NGF on sperm function.

机构信息

College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, Jilin Province, 130062, China.

出版信息

Theriogenology. 2010 Dec;74(9):1615-22. doi: 10.1016/j.theriogenology.2010.06.033. Epub 2010 Aug 12.

Abstract

The objective was to confirm the presence of nerve growth factor (NGF) and its specific receptor, TrkA, in ejaculated bovine sperm, and to investigate the effects of NGF on specific aspects of bovine sperm function. Both TrkA transcripts and immunoreactivity typical of the translated protein were detected by RT-PCR and western blotting. However, only the NGF protein was detected in bovine sperm using western blotting, and there was no RT-PCR evidence for NGF transcripts in sperm. Using an immunofluorescent technique, NGF-immunoreactivity was localized to the sperm head and tail, whereas that of TrkA was detected in the acrosomal cap, nucleus, and tail regions When sperm were treated with exogenous NGF, both leptin secretion and sperm viability were increased (P < 0.05); moreover, the percentages of late apoptotic and dead sperm were increased (P < 0.05). However, NGF had no effects on insulin secretion, mitochondrial activity, intracellular calcium levels, or the acrosome reaction of sperm (P > 0.05). In conclusion, the presence of TrkA transcript, as well as NGF and TrkA immunoreactivity were confirmed in bovine sperm. Furthermore, exogenous NGF had significant effects on the secretion of leptin, cell viability, and sperm apoptosis. This study provided strong evidence that NGF/TrkA may have roles in regulation of sperm physiology and perhaps male fertility and infertility.

摘要

目的是确认神经生长因子(NGF)及其特异性受体 TrkA 在射出牛精子中的存在,并研究 NGF 对牛精子功能特定方面的影响。通过 RT-PCR 和 Western blot 检测到 TrkA 转录本和翻译蛋白的典型免疫反应性。然而,Western blot 仅检测到牛精子中的 NGF 蛋白,而精子中没有 NGF 转录本的 RT-PCR 证据。使用免疫荧光技术,NGF-免疫反应性定位于精子头部和尾部,而 TrkA 则定位于顶体帽、核和尾部区域。当精子用外源性 NGF 处理时,瘦素分泌和精子活力均增加(P < 0.05);此外,晚期凋亡和死亡精子的百分比增加(P < 0.05)。然而,NGF 对胰岛素分泌、线粒体活性、细胞内钙水平或精子顶体反应没有影响(P > 0.05)。总之,在牛精子中证实了 TrkA 转录本以及 NGF 和 TrkA 免疫反应性的存在。此外,外源性 NGF 对瘦素分泌、细胞活力和精子凋亡有显著影响。本研究提供了强有力的证据表明,NGF/TrkA 可能在调节精子生理功能以及男性生育和不育方面发挥作用。

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