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小鼠卵母细胞激活过程中不伴随细胞内pH变化。

Intracellular pH change does not accompany egg activation in the mouse.

作者信息

Phillips K P, Baltz J M

机构信息

Loeb Medical Research Institute, Ottawa Civic Hospital, Ontario, Canada.

出版信息

Mol Reprod Dev. 1996 Sep;45(1):52-60. doi: 10.1002/(SICI)1098-2795(199609)45:1<52::AID-MRD8>3.0.CO;2-R.

Abstract

In the sea urchin, some other marine invertebrates, and the frog, Xenopus, egg activation at fertilization is accompanied by an increase in intracellular pH (pHi). We measured pHi in germinal vesicle (GV)-intact mouse oocytes, ovulated eggs, and in vivo fertilized zygotes using the pH indicator dye, SNARF-1. The mean pH, was 6.96 +/- 0.004 (+/- SEM) in GV-intact oocytes, 7.00 +/- 0.01 in ovulated, unfertilized eggs, and 7.02 +/- 0.01 in fertilized zygotes, indicating no sustained changes in pHi after germinal vesicle breakdown (GVBD) or fertilization. To examine whether transient changes in pHi occur shortly after egg activation, mouse eggs were parthenogenetically activated by 7% ethanol in phosphate buffered saline (PBS); no significant change in pHi followed ethanol activation. Since increased Na+/H+ antiporter activity is responsible for pHi increase in the sea urchin, pHi was measured in the absence of added bicarbonate or CO2 (a condition under which the antiporter would be the only major pHi regulatory mechanism able to operate, since the others were bicarbonate-dependent) in GV-intact oocytes, ovulated eggs, and in vivo fertilized zygotes to determine whether a Na+/H+ antiporter was activated. There was no physiologically significant difference in pHi after GVBD or fertilization, when pHi was measured in bicarbonate-free medium, nor any change upon parthenogenetic activation. Thus, a change in pHi is not a feature of egg activation in the mouse.

摘要

在海胆、一些其他海洋无脊椎动物以及青蛙(非洲爪蟾)中,受精时的卵子激活伴随着细胞内pH值(pHi)的升高。我们使用pH指示剂染料SNARF-1测量了生发泡(GV)完整的小鼠卵母细胞、排卵后的卵子以及体内受精的合子中的pHi。GV完整的卵母细胞中的平均pH值为6.96±0.004(±标准误),排卵后的未受精卵中的平均pH值为7.00±0.01,受精合子中的平均pH值为7.02±0.01,这表明在生发泡破裂(GVBD)或受精后pHi没有持续变化。为了研究卵子激活后不久pHi是否会发生短暂变化,小鼠卵子在磷酸盐缓冲盐水(PBS)中用7%乙醇进行孤雌激活;乙醇激活后pHi没有显著变化。由于海胆中Na+/H+反向转运蛋白活性增加是pHi升高的原因,因此在不添加碳酸氢盐或二氧化碳的情况下(在这种条件下,由于其他机制依赖碳酸氢盐,反向转运蛋白将是唯一能够起作用的主要pHi调节机制)测量了GV完整的卵母细胞、排卵后的卵子以及体内受精合子中的pHi,以确定Na+/H+反向转运蛋白是否被激活。当在无碳酸氢盐的培养基中测量时,GVBD或受精后pHi没有生理上的显著差异,孤雌激活后也没有任何变化。因此,pHi的变化不是小鼠卵子激活的特征。

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