Department of Biomedical and Molecular Sciences, Faculty of Health Sciences, Queen's University, Kingston, Ontario, K7L 3N6, Canada.
Yale Fertility Center, Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology & Reproductive Sciences, Yale University, Orange, Connecticut, 06477, USA.
J Assist Reprod Genet. 2022 Oct;39(10):2287-2301. doi: 10.1007/s10815-022-02591-0. Epub 2022 Aug 16.
To investigate the effects of recombinant human oviduct-specific glycoprotein (rHuOVGP1) alone and in combination with progesterone (P4) on intracellular Ca concentration [Ca] and to investigate if rHuOVGP1 in combination with P4 can further enhance tyrosine phosphorylation (pY) of sperm proteins during human sperm capacitation.
Fluorometric flow cytometry was performed to examine the effects of rHuOVGP1 on [Ca] in human sperm during capacitation. Confocal microscopy was used in conjunction with live cell imaging to analyze the influence of rHuOVGP1 and P4 on [Ca] in the sperm tail and to examine the involvement of CatSper channels in their effect on [Ca]. Western blot analysis was performed to assess the protein levels of p105, a major tyrosine-phosphorylated sperm protein.
rHuOVGP1 increases [Ca] in human sperm at the beginning of capacitation and further increases and sustains the level of [Ca] in the sperm tail following the addition of P4. Inhibition of CatSper channels impedes the effects of rHuOVGP1 on [Ca] in the sperm tail. P4 alone can increase pY of a major human sperm protein, p105, yet yields a further increase when used in combination with rHuOVGP1.
The present study revealed that rHuOVGP1 may work with P4 to upregulate [Ca] at the beginning of capacitation in part through CatSper channels which, in turn, leads to the downstream event of pY of sperm proteins and enhancement of sperm capacitation.
研究重组人输卵管特异性糖蛋白(rHuOVGP1)单独和与孕激素(P4)联合对细胞内 Ca 浓度[Ca]的影响,并研究 rHuOVGP1 与 P4 联合是否能进一步增强人精子获能过程中精子蛋白的酪氨酸磷酸化(pY)。
采用荧光流动细胞术检测 rHuOVGP1 在人精子获能过程中对[Ca]的影响。共聚焦显微镜与活细胞成像相结合,分析 rHuOVGP1 和 P4 对精子尾部[Ca]的影响,并研究 CatSper 通道在其对[Ca]影响中的作用。采用 Western blot 分析评估主要的酪氨酸磷酸化精子蛋白 p105 的蛋白水平。
rHuOVGP1 在人精子获能开始时增加[Ca],并在添加 P4 后进一步增加并维持精子尾部的[Ca]水平。CatSper 通道的抑制阻碍了 rHuOVGP1 对精子尾部[Ca]的影响。P4 单独可以增加主要的人精子蛋白 p105 的 pY,但与 rHuOVGP1 联合使用时会进一步增加。
本研究表明,rHuOVGP1 可能与 P4 一起在获能开始时上调[Ca],部分通过 CatSper 通道,进而导致精子蛋白 pY 的下游事件,并增强精子获能。