Protein tyrosine phosphorylation in boar sperm capacitation induced by a novel relaxin-like protein

新型松弛素样蛋白诱导公猪精子获能过程中的蛋白酪氨酸磷酸化

• 批准号: 18580280
• 负责人: KOHSAKA Tetsuya
• 金额: $ 2.57万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18580280/
• 关键词: Relaxin; Testis; Sperm; Capacitation; Phosphorvlation; Boar

This research project was planed to elucidate protein tyrosine phosphorylation during boar sperm capacitation and acrosome reaction mediated by relaxin-like protein (seminal relaxin). The outcome of this study as follows.The present study demonstrated that seminal relaxin actually induced boar sperm capacitation and acrosome reaction with high sperm viability. In relaxin-treated boar sperm, the activity of protein tyrosine kinase was increased with progress of the incubation time, which reached a plateau (10U/10^8 cells) at 4 h of incubation. Furthermore, Western blot analysis revealed a specific band of phosphorylated protein corresponding to the molecular size of 30 kDa in the capacitated/acrosome reacted boar sperm induced by relaxin, suggesting that relaxin induces tyrosine phosphorylation in boar sperm. Also proteome analysis characterized it as interesting and unusual. When polyclonal antibody was generated to the recombinant phosphorylated protein-MBP fusion protein expressed in E. coli, immunohistochemical analysis provided clear-cut evidence of the localization of phosphorylated protein in the sperm head and midpiece.

本研究旨在阐明猪精子获能过程中蛋白质酪氨酸的磷酸化以及由松弛素样蛋白(精液松弛素)介导的顶体反应。本研究的结果如下：本研究表明，精液松弛素能诱导猪精子获能和顶体反应，具有较高的精子存活率。在松弛素处理的猪精子中，蛋白酪氨酸激酶的活性随着孵育时间的延长而增加，在孵育4h时达到平台期(10U/10^8细胞)。此外，Western印迹分析显示，在松弛蛋白诱导的获能/顶体反应的猪精子中，有一条特定的磷酸化蛋白条带，对应于30 kDa的分子大小，这表明松弛蛋白诱导了猪精子中的酪氨酸磷酸化。此外，蛋白质组分析也将其描述为有趣和不寻常的。在大肠杆菌中表达的重组磷酸化蛋白-MBP融合蛋白产生多克隆抗体时，免疫组织化学分析为磷酸化蛋白在精子头部和中段的定位提供了明确的证据。

项目成果

Park EY Identification of epitope on DNA-binding protein expressed in insect cell infected by baculovirus

Park EY 杆状病毒感染昆虫细胞中表达的 DNA 结合蛋白表位的鉴定

• 发表时间: 2006
• 期刊: Molecular Biology Reports 33
• 作者: Suzuki;M.;Kohsaka;T
• 通讯作者: T

Effect of relaxin on motility, acrosome reaction and viability of cryopreserved boar spermatozoa

松弛素对冻存公猪精子运动、顶体反应和活力的影响

• 发表时间: 2006
• 期刊: Reproductive Medicine and Biology 5
• 作者: Suzuki;M.;Kohsaka;T;Miah A et al.
• 通讯作者: Miah A et al.

Identification of relaxin and its receptor LGR7 in the boar testis

公猪睾丸松弛素及其受体LGR7的鉴定

• 发表时间: 2006
• 期刊: Proceedings of 12th AAAP Animal Science Congress XII
• 作者: Maroyama;K.;Kato;S.;Aoshima;T.;Suchin;Itoh;H.;Yogo;K.;Kawarasaki;T.;Yamada;K.;Kitayama;Y.;Kohsaka;T
• 通讯作者: T

Effect of relaxin on the motility, acrosome reaction and utilization of glucose of fresh and frozen-thawed bovine spermatozoa

松弛素对新鲜和冻融牛精子运动、顶体反应和葡萄糖利用的影响

• 发表时间: 2007
• 期刊: Animal Science Journal 78
• 作者: Miah;AG.;Salma;U.;Tareq;K.;Kohsaka;T.;Tsuji;H
• 通讯作者: H

Expression and cellular pattern of relaxin mRNA in porcine corpora lutea during pregnancy

• DOI: 10.1007/s00441-007-0492-5
• 发表时间: 2007-11-01
• 期刊: CELL AND TISSUE RESEARCH
• 影响因子: 3.6
• 作者: Kohsaka, Tetsuya;Singh, Udai Pratap;Hashizume, Kazuyoshi
• 通讯作者: Hashizume, Kazuyoshi