Improvement of 5-HT<SUB>3</SUB> Receptor Binding Assay: Enhancement of Specific [<SUP>3</SUP>H]Quipazine Binding with Triton X-100-Treated Membranes from Rat Cortex
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概要
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The 5-hydroxytryptamine (5-HT)<SUB>3</SUB> receptor binding assay using [<SUP>3</SUP>H]quipazine was examined. It was impossible to obtain specific [<SUP>3</SUP>H]quipazine binding with the membrane fractions from rat cortex prepared by the usual procedure. When the membranes were pretreated with detergent Triton X-100, the ratio of specific [<SUP>3</SUP>H]quipazine binding markedly increased, depending upon the concentration of Triton X-100 in the range of 0.01-0.1% (w/v). At a concentration of more than 0.05%, the specific binding reached a maximum of 55 to 60% of the total binding. The specific [<SUP>3</SUP>H]quipazine binding to the Triton X-100-treated membranes was reversible and was potently inhibited by several 5-HT<SUB>3</SUB> antagonists, while 5-HT<SUB>1</SUB>, 5-HT<SUB>2</SUB> receptor antagonists and other receptor-specific ligands had no effect on the binding. Scatchard analysis indicated a single class of binding sites with a K<SUB>d</SUB> of 0.62 nM and B<SUB>max</SUB> of 97 fmol/mg protein. Thus, the Triton X-100-treated membranes retained the characteristics of 5-HT<SUB>3</SUB> binding sites, making it possible to use [<SUP>3</SUP>H]quipazine for a 5-HT<SUB>3</SUB> receptor binding assay with a high ratio of specific binding.
著者
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Furukawa Kiyoshi
Department Of Aquatic Bioscience Graduate School Of Agricultural And Life Sciences The University Of
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Komiya Masanobu
Department Of Pharmacology 1 Discovery Research Laboratories 1 Dainippon Pharmaceutical Co. Ltd.
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Furukawa Kiyoshi
Department of Pharmacology, Research Laboratories, Dainippon Pharmaceutical Co., Ltd.
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Une Teruaki
Department of Pharmacology, Research Laboratories, Dainippon Pharmaceutical Co., Ltd.
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