Expression of muscarinic M3-receptors coupled to inositol phospholipid hydrolysis in human detrusor cultured smooth muscle cells.
OBJECTIVE
To investigate the effect of muscarinic receptor agonists and antagonists on the accumulation of inositol phosphates in cultures of human detrusor smooth muscle cells.


METHODS
Primary explant culture was used to derive smooth muscle cell lines from small bladder biopsies. The cells were loaded with [3H]-myoinositol, stimulated with muscarinic agonists, and the accumulation of [3H]-inositol phosphates was measured by liquid scintillation counting.


RESULTS
Carbachol (EC50 8.3 microM.), methacholine (EC50 7.5 microM.), oxotremorine (EC50 2.5 microM.) and pilocarpine (EC50 8.3 microM.) produced concentration-dependent rises in the accumulation of total [3H]-inositol phosphates. M1 (pirenzepine), M2 (methoctramine) and M3 (4-DAMP and pf-HHSiD) muscarinic receptor antagonists significantly antagonized the response induced by a submaximal concentration of carbachol (100 microM.). The apparent pA2 values were atropine (9.4), 4-DAMP (9.2), pfHHSid (7.4), pirenzepine (6.9) and methoctramine (6.3).


CONCLUSIONS
These results indicate that human detrusor smooth muscle cells in culture express M3 muscarinic receptors which are linked to phosphoinositide hydrolysis.
Harriss
D R
DR
Department of Physiology and Pharmacology, Medical School, Queen's Medical Centre, Nottingham, United Kingdom.
Marsh
K A
KA
Birmingham
A T
AT
Hill
S J
SJ
D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D003959:Diamines / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D007295:Inositol Phosphates / Q000378:metabolism; D016210:Methacholine Chloride / Q000494:pharmacology; D018721:Muscarinic Agonists / Q000494:pharmacology; D009130:Muscle, Smooth / Q000378:metabolism; D010095:Oxotremorine / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000378:metabolism*; D010862:Pilocarpine / Q000494:pharmacology; D010890:Pirenzepine / Q000494:pharmacology; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000502:physiology*; D001743:Urinary Bladder / Q000378:metabolism*
D003959:Diamines; D007295:Inositol Phosphates; D018721:Muscarinic Agonists; D010716:Phosphatidylinositols; D011976:Receptors, Muscarinic; D010862:Pilocarpine; D016210:Methacholine Chloride; D010890:Pirenzepine; D010095:Oxotremorine; D002217:Carbachol; C054938:methoctramine