Analysis of Fertilization Phenomenon Using the Patch Clamp Method.

使用膜片钳方法分析受精现象。

基本信息

  • 批准号:
    01480404
  • 负责人:
  • 金额:
    $ 3.97万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
  • 财政年份:
    1989
  • 资助国家:
    日本
  • 起止时间:
    1989 至 1990
  • 项目状态:
    已结题

项目摘要

This research aimed at the analysis of fertilization phenomenon using the patch clamp method which is newly developed method in electro-physiology area.(1) Study of golden hamster ovumWe injected PMS and hCG in subcutaneous tissue of golden hamster. Hyperstimulated ova were collected using microscope and incubated in trypsin and hyaronidaze contained solution. Collected zona free ovum was attached with patch clamp electrode, and on-off phenomena of calcium ion channel were recorded and analyzed. After study of calcium ion channel in sperm free solution, sperm of golden hamster were added into solution. Frequency of on-off phenomena of calcium ion channel were increased after penetration of sperm into ovum under phase contrast microscope. These data are already speculated in other methods, but it is important to get these data in direct method.(2) Study of human ovumBecause studies of fertilization of human ovum are restricted under Japanese Obstetrics & gynecology Association now, the precise studies of human ovum cannot be done. We examined human ova which were collected in gynecological operations under permission of patients. Zona free ovum which was obtained after treatment of trypsin and hyaronidaze was attached with patch clamp electrode. Frequency of on-off phenomena of calcium ion channel was low in follicular phase, but in luteal phase it was high frequency. These data are newly found out.
本研究旨在利用电生理学领域新发展的膜片钳法对受精现象进行分析。(1)金黄地鼠卵的研究在金黄地鼠的皮下组织中注射PMS和hCG。使用显微镜收集过度刺激的卵子并在含有胰蛋白酶和透明质酸的溶液中孵育。将收集的无带卵子贴上膜片钳电极,记录并分析钙离子通道的开关现象。在研究了无精子溶液中的钙离子通道后,将金黄地鼠的精子添加到溶液中。相差显微镜下精子穿入卵子后钙离子通道开关现象的频率增加。这些数据已经用其他方法推测出来,但用直接法获得这些数据很重要。(2)人类卵子的研究由于人类卵子受精的研究目前受到日本妇产科学会的限制,因此无法对人类卵子进行精确的研究。我们检查了在患者许可下在妇科手术中收集的人类卵子。将胰蛋白酶和透明酶处理后获得的无带卵用膜片钳电极附着。钙离子通道开关现象在卵泡期频率较低,但在黄体期频率较高。这些数据是新发现的。

项目成果

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