Quantifying receptor properties: the tissue segment binding method - a powerful tool for the pharmacome analysis of native receptors.
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.
Muramatsu
Ikunobu
I
Division of Pharmacology, Department of Biochemistry and Bioinformative Sciences, School of Medicine, University of Fukui, Matsuoka, Fukui 910-1193, Japan. muramatu@fmsrsa.fukui-med.ac.jp
Tanaka
Takashi
T
Suzuki
Fumiko
F
Li
Zhang
Z
Hiraizumi-Hiraoka
Yasuko
Y
Anisuzzaman
Abu Syed Md
AS
Yamamoto
Hatsumi
H
Horinouchi
Takahiro
T
Morishima
Shigeru
S
D000818:Animals; D001667:Binding, Competitive; D006801:Humans; D008954:Models, Biological; D011869:Radioligand Assay / Q000379:methods*; D011956:Receptors, Cell Surface / Q000032:analysis*
D011956:Receptors, Cell Surface