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导致海星卵激活的酪氨酸激酶和G蛋白偶联途径的证据。

Evidence for both tyrosine kinase and G-protein-coupled pathways leading to starfish egg activation.

作者信息

Shilling F M, Carroll D J, Muslin A J, Escobedo J A, Williams L T, Jaffe L A

机构信息

Department of Physiology, University of Connecticut Health Center, Farmington 06032.

出版信息

Dev Biol. 1994 Apr;162(2):590-9. doi: 10.1006/dbio.1994.1112.

Abstract

To investigate possible pathways leading to egg activation at fertilization, the ability of exogenously introduced tyrosine kinase and G-protein-coupled receptors to mimic events of fertilization was examined. Oocytes of the starfish Asterina miniata were injected with RNA for a chimeric receptor consisting of the extracellular domain of the beta form of the mouse platelet-derived growth factor (PDGF) receptor and the transmembrane/intracellular domain of the human fibroblast growth factor (FGF) receptor, or with RNA for the rat serotonin 1c receptor. These oocytes were cultured for 1 to 3 days and then matured with 1-methyladenine. In response to PDGF or serotonin, the injected eggs underwent responses like those at fertilization: cortical granule exocytosis, a rise in intracellular free calcium, and DNA synthesis. Some of these artificially activated eggs cleaved, and some of the PDGF-activated eggs were observed to form larvae. A PDGF/FGF receptor with a point mutation which eliminated its ability to interact with phospholipase C-gamma did not cause fertilization-like responses. Thus components of a signaling pathway involving phospholipase C-gamma, characteristic of tyrosine kinase receptors, as well as components of a pathway involving a G-protein and phospholipase C-beta, characteristic of G-protein-coupled receptors, appear to be present in starfish eggs. Either or both could function in egg activation at fertilization.

摘要

为了研究受精时导致卵母细胞激活的可能途径,研究人员检测了外源性引入的酪氨酸激酶和G蛋白偶联受体模拟受精事件的能力。将由小鼠血小板衍生生长因子(PDGF)受体β形式的细胞外结构域与人成纤维细胞生长因子(FGF)受体的跨膜/细胞内结构域组成的嵌合受体的RNA,或大鼠5-羟色胺1c受体的RNA注射到小海星(Asterina miniata)的卵母细胞中。将这些卵母细胞培养1至3天,然后用1-甲基腺嘌呤使其成熟。响应PDGF或5-羟色胺,注射后的卵母细胞会出现类似受精时的反应:皮质颗粒胞吐、细胞内游离钙升高和DNA合成。一些人工激活的卵母细胞发生了分裂,并且观察到一些PDGF激活的卵母细胞形成了幼虫。具有点突变且消除了其与磷脂酶C-γ相互作用能力的PDGF/FGF受体不会引起类似受精的反应。因此,海星卵母细胞中似乎存在涉及磷脂酶C-γ的信号通路成分(酪氨酸激酶受体的特征)以及涉及G蛋白和磷脂酶C-β的信号通路成分(G蛋白偶联受体的特征)。其中任何一个或两者都可能在受精时的卵母细胞激活中发挥作用。

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