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使用PC12细胞对调节性分泌进行分析。

Analysis of regulated secretion using PC12 cells.

作者信息

Taupenot Laurent

机构信息

University of California at San Diego, La Jolla, California, USA.

出版信息

Curr Protoc Cell Biol. 2007 Sep;Chapter 15:Unit 15.12. doi: 10.1002/0471143030.cb1512s36.

Abstract

The catecholamine-secreting PC12 cell line derived from the rat adrenal medulla has long been considered a model system for neurosecretion and neuronal differentiation. PC12 cells contain a large number of secretory granules (otherwise known as large dense-core vesicles) for storage of small molecules, processing enzymes, neuropeptides, and peptide hormones. Secretory granule exocytosis in PC12 cells is tightly regulated by calcium and occurs in response to a secretagogue. This unit provides protocols for maintenance and transfection of PC12 cells. Several secretion assays are described to measure the release of secretory granule cargo molecules by detection of radioactive catecholamine, or by immunochemical or chemiluminescence detection of transfected regulated secretory proteins.

摘要

源自大鼠肾上腺髓质的分泌儿茶酚胺的PC12细胞系长期以来一直被视为神经分泌和神经元分化的模型系统。PC12细胞含有大量分泌颗粒(也称为大致密核心囊泡),用于储存小分子、加工酶、神经肽和肽激素。PC12细胞中的分泌颗粒胞吐作用受钙的严格调控,并在促分泌素的作用下发生。本单元提供了PC12细胞的培养和转染方案。描述了几种分泌测定方法,通过检测放射性儿茶酚胺,或通过免疫化学或化学发光检测转染的调节性分泌蛋白,来测量分泌颗粒货物分子的释放。

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