Muramatsu Ikunobu, Tanaka Takashi, Suzuki Fumiko, Li Zhang, Hiraizumi-Hiraoka Yasuko, Anisuzzaman Abu Syed Md, Yamamoto Hatsumi, Horinouchi Takahiro, Morishima Shigeru
Division of Pharmacology, Department of Biochemistry and Bioinformative Sciences, School of Medicine, University of Fukui, Matsuoka, Fukui 910-1193, Japan.
J Pharmacol Sci. 2005 Aug;98(4):331-9. doi: 10.1254/jphs.cpj05001x. Epub 2005 Aug 5.
The radioligand binding assay technique is an extremely powerful tool for studying receptors. It allows an analysis of the interactions of hormones, neurotransmitters, and related drugs with their receptors. Most of the binding assays have widely been applied to crude membrane fractions prepared from many tissues, but in the conventional method, there are some limitations such as a yield loss of receptor-bearing membranes and a change in receptor environment upon homogenization and fractionation. Recently, in order to overcome these problems, a binding assay has been developed using intact tissue segments. This article presents a brief overview of the tissue segment binding assay that has been developed mainly in our department. Practical guidelines for setting up this new assay are presented, including segment preparation, choice of appropriate radioligand, optimizing assay conditions, and appropriate methods for data analysis. The unique advantages and disadvantages of the tissue segment binding method are discussed in comparison with those of conventional membrane binding methods. We suggest that the tissue segment binding method is a powerful tool for detecting the native properties of receptors occurring in tissues and cells without altering their environment.
放射性配体结合测定技术是研究受体的一种极其强大的工具。它能够分析激素、神经递质及相关药物与其受体的相互作用。大多数结合测定已广泛应用于从许多组织制备的粗制膜组分，但在传统方法中，存在一些局限性，如含受体膜的产量损失以及匀浆和分级分离时受体环境的变化。最近，为了克服这些问题，已开发出使用完整组织切片的结合测定法。本文简要概述了主要在我们科室开发的组织切片结合测定法。文中给出了建立这种新测定法的实用指南，包括切片制备、合适放射性配体的选择、测定条件的优化以及数据分析的适当方法。与传统膜结合方法相比，讨论了组织切片结合方法独特的优缺点。我们认为，组织切片结合方法是一种强大的工具，可用于检测组织和细胞中受体的天然特性，而不会改变其环境。
