Cholesterol efflux from the boar sperm for a novel relaxin-like protein and its action on their capacitation

公猪精子中的胆固醇流出，产生一种新型松弛素样蛋白及其对获能的作用

• 批准号: 13660283
• 负责人: KOHSAKA Tetsuya
• 金额: $ 2.3万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660283/
• 关键词: Relaxin; sperm; cholesterol efflux; Capacitation; Boar; コレステロール; アクセプター

This research project was planned to elucidate the physiological role of relaxin-like protein (RLP) during sperm capacitation in boars. In detail, I will 1) determine whether RLP stimulates cholesterol efflux from the plasma membrane of boar sperm during capacitation, 2) identify the sites of cholesterol release on boar spermbinding sites on boar sperm for relaxin-like protein, and 3) RLP potentiates sperm capacitating. The outcome of this research is as follows.1. Induction of sperm capacitation : RLP induced sperm capacitation and acrosome reaction in a dose-dependent manner, with maximum effect when 150μg/ml (11μM) RLP was added.2. Stimulation of cholesterol efflux : RLP stimulated cholesterol efflux from the plasma membrane on lboar sperm labeled with [^3H] cholesterol.3. Identification of the site(s) of cholesterol efflux : RLP stimulated cholesterol efflux from the acrosomal region of boar sperm, as revealed by the filipin-cholesterol staining.4. Possible physiological roles on boar sperm : RLP promotes capacitation by facilitating cholesterol efflux from the sperm plasma membrane covering the acrosomal region, suggesting that RLP is the most important factor in boar seminal plasma responsible for stimulation of sperm capacitation.

本研究旨在阐明松弛素样蛋白（RLP）在公猪精子获能过程中的生理作用。详细地说，我将1)确定RLP是否在获能过程中刺激胆固醇从猪精子的质膜流出，2)确定在猪精子上的松弛素样蛋白结合位点上释放胆固醇的位点，以及3)RLP增强精子的获能。本研究的结果如下：1。诱导精子获能：RLP诱导精子获能和顶体反应呈剂量依赖性，当RLP浓度为150μg/ml （11μM）时效果最大。2 .刺激胆固醇外排：RLP刺激[^3H]胆固醇标记的猪精子的质膜上的胆固醇外排。胆固醇外排部位的鉴定：菲律宾胆固醇染色显示，RLP刺激了猪精子顶体区域的胆固醇外排。对猪精子可能的生理作用：RLP通过促进胆固醇从覆盖顶体区域的精子质膜流出来促进获能，这表明RLP是猪精浆中负责刺激精子获能的最重要因素。

项目成果

T. Kohsaka et al.: "Ultrastractural properties and immunolocalization of relaxin in the cvtoplasmic electron-dense granules of large luteal cells during pregnancy"Journal of Reproduction and Development. 47. 217-225 (2001)

T. Kohsaka 等人：“怀孕期间大黄体细胞胞质电子致密颗粒中松弛素的超结构特性和免疫定位”生殖与发育杂志。

T.Kohsaka et al.: "Ultrastructural properties and immunolocalization of relaxin in the cytoplasmic electron-dense granules of large luteal cells during pregnancy in the cow"Journal of Reproduction and Fertility. 47. 217-225 (2001)

T.Kohsaka 等人：“牛怀孕期间大黄体细胞细胞质电子致密颗粒中松弛素的超微结构特性和免疫定位”生殖与生育杂志。

T.Kohsaka et al.: "Seminal Immunoreactive Relaxin in Domestic Animals and Its Relationship to Sperm Motility as a Possible Index for Predicting the Fertilizing Ability of Sires"International Journal of Andrology. 26. 115-120 (2003)

T.Kohsaka 等人：“家畜精液免疫反应性松弛素及其与精子活力的关系，作为预测雄性受精能力的可能指标”国际男科杂志。

J.Kobayashi et al.: "Effect of lactoferrin on motility and capacitation of bovine spermatozoa"Theriogenology. 59. 46 (2003)

J.Kobayashi 等人：“乳铁蛋白对牛精子活力和获能的影响”动物发生学。

T. Kohsaka et al.: "The presence of specific binding sites on boar spermatozoa for porcine relaxin and its action on their motility characteristics"Journal of Reproduction and Development. 47. 197-204 (2001)

T. Kohsaka 等人：“公猪精子上猪松弛素特异性结合位点的存在及其对其运动特性的作用”《生殖与发育杂志》。