Carbachol regulates pacemaker activities in cultured interstitial cells of Cajal from the mouse small intestine.
We studied the effect of carbachol on pacemaker currents in cultured interstitial cells of Cajal (ICC) from the mouse small intestine by muscarinic stimulation using a whole cell patch clamp technique and Ca2+-imaging. ICC generated periodic pacemaker potentials in the current-clamp mode and generated spontaneous inward pacemaker currents at a holding potential of-70 mV. Exposure to carbachol depolarized the membrane and produced tonic inward pacemaker currents with a decrease in the frequency and amplitude of the pacemaker currents. The effects of carbachol were blocked by 1-dimethyl-4-diphenylacetoxypiperidinium, a muscarinic M(3) receptor antagonist, but not by methotramine, a muscarinic M(2) receptor antagonist. Intracellular GDP-beta-S suppressed the carbachol-induced effects. Carbachol-induced effects were blocked by external Na+-free solution and by flufenamic acid, a non-selective cation channel blocker, and in the presence of thapsigargin, a Ca2+-ATPase inhibitor in the endoplasmic reticulum. However, carbachol still produced tonic inward pacemaker currents with the removal of external Ca2+. In recording of intracellular Ca2+ concentrations using fluo 3-AM dye, carbachol increased intracellular Ca2+ concentrations with increasing of Ca2+ oscillations. These results suggest that carbachol modulates the pacemaker activity of ICC through the activation of non-selective cation channels via muscarinic M(3) receptors by a G-protein dependent intracellular Ca2+ release mechanism.
So
Keum Young
KY
Department of Anesthesiology, College of Medicine, Chosun University, Gwangju 501-759, Korea.
Kim
Sang Hun
SH
Sohn
Hong Moon
HM
Choi
Soo Jin
SJ
Parajuli
Shankar Prasad
SP
Choi
Seok
S
Yeum
Cheol Ho
CH
Yoon
Pyung Jin
PJ
Jun
Jae Yeoul
JY
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D001683:Biological Clocks / Q000187:drug effects* / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002217:Carbachol / Q000494:pharmacology*; D027682:Cation Transport Proteins / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D003959:Diamines / Q000494:pharmacology; D005439:Flufenamic Acid / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D007421:Intestine, Small / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D009467:Neuromuscular Depolarizing Agents / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D010880:Piperidines / Q000494:pharmacology*; D018517:Plant Roots; D043585:Receptor, Muscarinic M2 / Q000037:antagonists & inhibitors; D043587:Receptor, Muscarinic M3 / Q000037:antagonists & inhibitors*; D053629:Thapsia; D019284:Thapsigargin / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology
D000893:Anti-Inflammatory Agents; D027682:Cation Transport Proteins; D003959:Diamines; D009467:Neuromuscular Depolarizing Agents; D010880:Piperidines; D043585:Receptor, Muscarinic M2; D043587:Receptor, Muscarinic M3; D013873:Thionucleotides; D006153:Guanosine Diphosphate; D005439:Flufenamic Acid; D019284:Thapsigargin; C023427:guanosine 5'-O-(2-thiodiphosphate); C042375:4-diphenylacetoxy-1,1-dimethylpiperidinium; D002217:Carbachol; D000252:Calcium-Transporting ATPases; C054938:methoctramine; D002118:Calcium