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HIGH‐LEVEL EXPRESSION OF RAT D 1A DOPAMINE RECEPTOR cDNA IN MOUSE FIBROBLAST LTK‐ CELLS BY n ‐BUTYRATE
Author(s) -
Horiuchi Akira,
Felder Robin A.
Publication year - 1996
Publication title -
clinical and experimental pharmacology and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.752
H-Index - 103
eISSN - 1440-1681
pISSN - 0305-1870
DOI - 10.1111/j.1440-1681.1996.tb02587.x
Subject(s) - adenylyl cyclase , biology , receptor , medicine , butyrate , endocrinology , microbiology and biotechnology , agonist , dopamine receptor , receptor expression , complementary dna , receptor antagonist , dopamine , antagonist , biochemistry , gene , fermentation
SUMMARY 1. In order to develop a simple, efficient system for the high‐level expression of dopamine receptors in eukaryotic cells, we have studied the effects of n ‐butyrate on the expression of rat D 1A dopamine receptor cDNA in mouse fibroblast LTK ‐ cells as compared with those of n ‐butyrate on endogenous D 1 receptor levels in opossum kidney cells. 2. In the transfected LTK ‐ cell membranes with pRc/CMV‐D 1A receptor cDNA, a selective D 1 dopamine antagonist, [ 3 H]‐SCH 23390, exhibited a K 4 of 0.9 ± 0.1 nmol/L and a B max of 0.35 ± 0.05 pmol/mg protein ( n = 5). 3. Addition of n ‐butyrate (2–10 mmol/L) to the culture medium for 48 h dose‐dependently increased the D 1A receptor level up to 1.5 ± 0.3 pmol/mg protein ( n = 7), although the K 4 values were not affected. The increase in receptor level was accompanied by an elevation of selective D 1 agonist‐induced adenylyl cyclase activity. 4. In contrast, n ‐butyrate treatment (2–10 mmol/L) did not affect either endogenous D 1 receptor levels or fendoldopam‐induced adenylyl cyclase activity in opossum kidney cells. 5. These results suggest n ‐butyrate is a useful tool for obtaining high‐level expression of D 1A dopamine receptor cDNA in mouse fibroblast LTK ‐ cells.