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钙调蛋白参与了用环腺苷酸类似物孵育的猪射出精液中细胞外 Ca 依赖性完全型超激活的发生。

Calmodulin is involved in the occurrence of extracellular Ca -dependent full-type hyperactivation in boar ejaculated spermatozoa incubated with cyclic AMP analogs.

机构信息

Laboratory of Reproductive Biology, Division of Animal Science, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, Kobe, Japan.

出版信息

Anim Sci J. 2021 Jan-Dec;92(1):e13552. doi: 10.1111/asj.13552.

Abstract

In mammals, hyperactivation is essential for sperm fertilization with oocytes in vivo. Two types of hyperactivation "full-type and nonfull-type patterns" can be observed in the spermatozoa from boars, bulls, and mice. We have a hypothesis that the full-type hyperactivation is a physiological (in vivo) pattern and are elucidating its molecular bases. The aims of this study were to detect calmodulin in boar sperm flagella by Western blotting and indirect immunofluorescence and to investigate effects of extracellular Ca and calmodulin antagonists "W-7 and W-5 (W-5; a less potent antagonist)" on the occurrence of full-type hyperactivation in boar spermatozoa. Calmodulin was specifically detected as the 17-kDa antigen in the flagella and postacrosomal region of the heads. Full-type hyperactivation could be induced effectively in the samples incubated with 3.42 mM CaCl for 120-180 min, and it was significantly reduced in the concentration-dependent manners of W-7 and W-5. Suppressing effects of W-7 on the full-type hyperactivation were stronger than those of W-5. These observations indicate that flagellar calmodulin is involved in the occurrence of extracellular Ca -dependent full-type hyperactivation in boar spermatozoa. This is the first indication of the intracellular Ca -sensing molecule which can function in the full-type hyperactivation.

摘要

在哺乳动物中,精子在体内与卵子受精时必须超激活。在猪、牛和老鼠的精子中可以观察到两种类型的超激活“全型和非全型模式”。我们假设全型超激活是一种生理(体内)模式,并正在阐明其分子基础。本研究的目的是通过 Western 印迹和间接免疫荧光检测猪精子鞭毛中的钙调蛋白,并研究细胞外 Ca 和钙调蛋白拮抗剂“W-7 和 W-5(W-5;一种效力较弱的拮抗剂)”对猪精子全型超激活发生的影响。钙调蛋白在鞭毛和头部的顶体后区特异性地被检测为 17-kDa 抗原。在 3.42 mM CaCl 孵育 120-180 分钟的样品中,可以有效地诱导全型超激活,并且 W-7 和 W-5 的浓度依赖性方式显著降低。W-7 对全型超激活的抑制作用强于 W-5。这些观察结果表明,鞭毛钙调蛋白参与了猪精子中细胞外 Ca 依赖性全型超激活的发生。这是第一个可以在全型超激活中发挥作用的细胞内 Ca 感应分子的迹象。

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