High-level expression of rat D1A dopamine receptor cDNA in mouse fibroblast LTK- cells by n-butyrate.
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 D1A dopamine receptor cDNA in mouse fibroblast LTK- cells as compared with those of n-butyrate on endogenous D1 receptor levels in opossum kidney cells. 2. In the transfected LTK- cell membranes with pRc/CMV-D1A receptor cDNA, a selective D1 dopamine antagonist, [3H]-SCH 23390, exhibited a Kd of 0.9 +/- 0.1 nmol/L and a Bmax 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 D1A receptor level up to 1.5 +/- 0.3 pmol/mg protein (n = 7), although the Kd values were not affected. The increase in receptor level was accompanied by an elevation of selective D1 agonist-induced adenylyl cyclase activity. 4. In contrast, n-butyrate treatment (2-10 mmol/L) did not affect either endogenous D1 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 D1A dopamine receptor cDNA in mouse fibroblast LTK- cells.
Horiuchi
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A
The Second Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan.
Felder
R A
RA
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001552:Benzazepines / Q000378:metabolism*; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D002460:Cell Line; D018076:DNA, Complementary / Q000187:drug effects / Q000378:metabolism*; D018492:Dopamine Antagonists / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D009893:Opossums; D051381:Rats; D011954:Receptors, Dopamine / Q000187:drug effects / Q000378:metabolism*
D001552:Benzazepines; D002087:Butyrates; D018076:DNA, Complementary; D018492:Dopamine Antagonists; D011954:Receptors, Dopamine; D020148:Butyric Acid; D000262:Adenylyl Cyclases