The cloned murine M1 muscarinic receptor is associated with the hydrolysis of phosphatidylinositols in transfected murine B82 cells.
A rat genomic DNA clone was isolated by its homology with a conserved primary sequence among the mammalian and avian beta adrenergic and porcine muscarinic receptors. A gene identified in this clone was highly homologous to the rat M1 muscarinic receptor. Stable expression of this gene was achieved in an established murine fibroblast cell line, B82. The gene product exhibits M1 type muscarinic receptor characteristics, as it has high affinity for PZ but low affinity for AF-DX 116. Carbachol stimulated the hydrolysis of phosphatidylinositols in the transfected cells. Pirenzepine had a more potent inhibitory effect on this response than AF-DX 116 since their functional inhibition constants were 13 nM and 480 nM, respectively, which is consistent with an M1 pharmacological profile. These data suggest that the M1 muscarinic receptor encoded by the gene is coupled to the hydrolysis of phosphatidylinositols after transfecting this gene into the B82 cells.
Lai
J
J
Section of Receptor Biochemistry and Molecular Biology, National Institute of Neurologic, Communicative Disorders and Stroke, Bethesda, MD 20892.
Mei
L
L
Roeske
W R
WR
Chung
F Z
FZ
Yamamura
H I
HI
Venter
J C
JC
D000818:Animals; D001285:Atropine / Q000494:pharmacology; D001667:Binding, Competitive; D002217:Carbachol / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004274:DNA, Recombinant; D005347:Fibroblasts / Q000378:metabolism*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D006868:Hydrolysis; D051379:Mice; D009116:Muscarine / Q000037:antagonists & inhibitors; D009693:Nucleic Acid Hybridization; D010716:Phosphatidylinositols / Q000378:metabolism*; D010890:Pirenzepine / Q000031:analogs & derivatives / Q000494:pharmacology; D011813:Quinuclidinyl Benzilate / Q000378:metabolism; D051381:Rats; D011976:Receptors, Muscarinic / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D004274:DNA, Recombinant; D010716:Phosphatidylinositols; D011976:Receptors, Muscarinic; D011994:Recombinant Proteins; D006165:Guanylyl Imidodiphosphate; D010890:Pirenzepine; D011813:Quinuclidinyl Benzilate; D001285:Atropine; D009116:Muscarine; D002217:Carbachol; C048593:otenzepad