High-level stable expression of recombinant 5-HT1A 5-hydroxytryptamine receptors in Chinese hamster ovary cells
| Autor: | R Wootton, P G Strange, A Newman-Tancredi |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Tetrahydronaphthalenes
Growth-hormone-releasing hormone receptor Receptor expression B-cell receptor Gene Expression CHO Cells Biology Transfection Binding Competitive Hippocampus Biochemistry Beta-1 adrenergic receptor Cricetinae Muscarinic acetylcholine receptor M5 Animals Humans heterocyclic compounds 5-HT5A receptor Molecular Biology 8-Hydroxy-2-(di-n-propylamino)tetralin Chinese hamster ovary cell Cell Membrane Colforsin Cell Biology Blotting Northern Molecular biology Recombinant Proteins Receptors Serotonin Cattle Guanosine Triphosphate Cyclase activity Adenylyl Cyclases Research Article |
| Zdroj: | Scopus-Elsevier |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj2850933 |
| Popis: | The human 5-hydroxytryptamine 5-HT1A receptor gene was transfected into Chinese hamster ovary cells. A series of recombinant monoclonal cell lines expressing the receptor were isolated and the properties of one cell line that expressed receptors at a high level (2.8 pmol/mg) were studied in detail. In ligand binding assays with the selective 5-HT1A receptor agonist 2-(NN-di[3H]propylamino)-8-hydroxy-1,2,3,4-tetrahydronaphthalene ([3H]8-OH-DPAT) only a single class of saturable high-affinity binding sites was detected, with a pharmacological profile in competition experiments essentially identical to that of the 5-HT1A receptor of bovine hippocampus. [3H]8-OH-DPAT binding to the recombinant cell membranes was inhibited by GTP, showing that the receptors in the transfected cells couple to G-proteins. A series of 5-hydroxytryptamine agonists inhibited forskolin-stimulated adenylate cyclase activity in the cells and, despite the high level of receptor expression, their apparent efficacies were similar to those observed for inhibition of adenylate cyclase in brain. This recombinant cell line provides a complete model system for studying the 5-HT1A receptor and its transmembrane signalling system. The recombinant cells can also be grown in suspension culture for long periods but, whereas 5-HT1A receptor numbers and receptor regulation by guanine nucleotides are maintained in suspension-grown cells, the inhibition of adenylate cyclase by the 5-HT1A receptor is gradually lost. |
| Databáze: | OpenAIRE |
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