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整合素介导小鼠卵内钙离子增加

岳利民, 张蕾, 何亚平, 张金虎, 谢怡敏, 郑煜, 张立, 黄萍, 王丽, 刘伟信

四川大学华西基础医学与法医学院生理学教研室. 四川, 成都 610041；四川大学华西医院. 四川, 成都 610041；成都市计划生育指导所. 四川, 成都 610031

摘要

为了研究整合素是否作为跨膜信号传递受体介导小鼠卵[Ca~(2+)]_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[Ca~(2+)]_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[Ca~(2+)]_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[Ca~(2+)]_i增加,增加的程度与精子作用相似;去除培养液中的Ca~(2+)_i后,再用RGD肽、Fn作用仍可引起卵[Ca~(2+)]_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[Ca~(2+)]_i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[Ca~(2+)]_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[Ca~(2+)]_i增加。

关键词： 整合素; 小鼠卵; 钙离子; 酪氨酸激酶; 蛋白激酶C

Integrins mediate the increase of concentration of intracellular free calcium in mouse eggs

Yueli Min, Zhang Lei, He Yaping, Zhang Jinhu, Xie Yimin, Zheng Yu, Zhang Li, Huang Ping, Wang Li, Liu Weixin

Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan Univeristy. Chengdu 610041, China；West China Hospital, Sichuan University. Chengdu 610041, China；Chengdu Institute of Family Planning. Chendu 610015, China

Abstract

For studying the effect of integrin on the [Ca~(2+)]_i of mouse eggs and its transmembrane signaling mechanism,zona-free mouse eggs were loaded with calcium probe Fluo-3/AM and the intensity of fluorescence of the eggs treated with different factors was measured through laser confocal microscopy. The results showed that the [Ca~(2+)], of zona-free mouse eggs was increased when the eggs were treated with RGD peptide, fibronectin (Fn) and anti-mouse integrin subunit α_6 and β_1 monoclonal antibodies, respectively. The [Ca~(2+)]_i, of the mouse eggs was also increased when the eggs were placed in calcium-free medium and treated with RGD peptide or Fn. The changes in the mouse egg [Ca~(2+)] _i caused by RGD and Fn were similar to those caused by sperm.

Key words: Integrins;Mouse oocyte;Intracellular calcium;Tyrosine kinase;Protein kinase C

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引用本文：

岳利民, 张蕾, 何亚平, 张金虎, 谢怡敏, 郑煜, 张立, 黄萍, 王丽, 刘伟信. 整合素介导小鼠卵内钙离子增加[J]. 生理学报 2004; 56 (3): .

Yueli Min, Zhang Lei, He Yaping, Zhang Jinhu, Xie Yimin, Zheng Yu, Zhang Li, Huang Ping, Wang Li, Liu Weixin. Integrins mediate the increase of concentration of intracellular free calcium in mouse eggs. Acta Physiol Sin 2004; 56 (3): (in Chinese with English abstract).