Involvement of G alpha q/11 in the contractile signal transduction pathway of muscarinic M3 receptors in caecal smooth muscle.
The nature of the pertussis toxin-insensitive G-protein involved in muscarinic-mediated phosphoinositides breakdown and contraction of isolated smooth muscle cells from the circular layer of the rabbit caecum was investigated. Immunoblotting of membrane proteins using affinity purified antibodies directed against different G-protein alpha-subunits revealed the expression of G alpha q/11, G alpha 11 and G alpha 12 in these cells. The carbachol-mediated [3H]inositol phosphates accumulation in saponin-permeabilized cells was abolished by anti-G alpha q/11-antibodies whereas anti-G alpha i1,2-antibodies were ineffective. Moreover, the carbachol-induced contraction of permeabilized cells, as determined by videomicrocopic measurements, was reversed by anti-G alpha q/11-antibodies but not affected by anti-G alpha i1,2-antibodies. From these data, we conclude that carbachol stimulates phosphoinositides hydrolysis and cell contraction through activation of specific muscarinic M3 receptors coupled to the pertussis toxin-insensitive G alpha q/11-protein. This is the first demonstration of G alpha q/11 implication in the contractile signal transduction pathway of muscarinic M3 receptors in smooth muscle cells.
Cuq
P
P
Laboratoire de Biochimie des Membranes, INSERM CJF 92-07, Université de Montpellier I. Faculté de Pharmacie, France.
Zumbihl
R
R
Fischer
T
T
Rouot
B
B
Bali
J P
JP
Magous
R
R
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002432:Cecum; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000502:physiology*; D008297:Male; D018721:Muscarinic Agonists / Q000494:pharmacology; D009119:Muscle Contraction / Q000502:physiology*; D009130:Muscle, Smooth / Q000187:drug effects; D011817:Rabbits; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D010738:Type C Phospholipases / Q000378:metabolism
D000906:Antibodies; D018721:Muscarinic Agonists; D011976:Receptors, Muscarinic; D002217:Carbachol; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins