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Integrins mediate the increase of concentration of intracellular free calcium in mouse eggs
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- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 3
- DOI:
- 10.3321/j.issn:0371-0874.2004.03.012
- Key Word:
- 整合素;小鼠卵;钙离子;酪氨酸激酶;蛋白激酶C
Abstract： 为了研究整合素是否作为跨膜信号传递受体介导小鼠卵[Ca2+]i的变化并探讨其机制.本实验采用甘-精-甘-天冬-丝-脯(GLY-ARG-GLY-ASP-SER-PRO,RGD肽)、纤连蛋白(fibronectin,Fn)及抗整合素α6、β1的单克隆抗体作用于负载了钙探针Fluo-3/AM的去透明带小鼠卵,用激光共聚焦显微镜检测小鼠卵的荧光强度以反映卵[Ca2+]i;用无钙液替代有钙液、或用酪氨酸激酶抑制剂或蛋白激酶C的抑制剂预先作用于卵,然后再观察RGD肽所致卵[Ca2+]i的变化.结果显示整合素配体RGD肽或Fn作用于去透明带小鼠卵可引起卵[Ca2+]i增加,增加的程度与精子作用相似;去除培养液中的Ca2+后,再用RGD肽、Fn作用仍可引起卵[Ca2+]i增加;用功能性的抗小鼠整合素α6、β1的单克隆抗体也可引起不同程度的卵[Ca2+]1增加,尤其以抗小鼠整合素α6单克隆抗体的作用明显;用酪氨酸激酶抑制剂预先作用于鼠卵,RGD肽或精子作用都不再引起卵[Ca2+]i增加;蛋白激酶C抑制剂预先作用鼠卵,RGD肽及Fn也不再引起卵[Ca2+]i增加.实验证明,小鼠卵膜整合素与其配体结合可使卵内贮存钙离子释放,引起卵[Ca2+]i增加这一卵激活的早期事件;整合素介导小鼠卵激活需要酪氨酸激酶信号转导途径的参与;蛋白激酶C也参与了整合素介导的卵激活.
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