Carbon monoxide activates human intestinal smooth muscle L-type Ca2+ channels through a nitric oxide-dependent mechanism.
Carbon monoxide (CO) is increasingly recognized as a physiological messenger. CO is produced in the gastrointestinal tract with diverse functions, including regulation of gastrointestinal motility, interacting with nitric oxide (NO) to mediate neurotransmission. The aim of this study was to determine the effect of CO on the human intestinal L-type Ca(2+) channel expressed in HEK cells and in native cells using the patch-clamp technique. Extracellular solution contained 10 mM Ba(2+) as the charge carrier. Maximal peak Ba(2+) current (I(Ba)) was significantly increased by bath application of 0.2% CO to transfected HEK cells (18 +/- 3%). The NO donor S-nitroso-N-acetylpenicillamine also increased I(Ba), and CO (0.2%) increased NO production in transfected HEK cells. The CO-induced increase in I(Ba) was blocked when cells were pretreated with 1H-[1,2,4]-oxadiazolo[4,3-a]quinoxalin-1-one (10 microM) or inhibitors of NO synthase (NOS). The PKA inhibitor KT-5720 (0.5 microM) and milrinone (3 microM), a phosphodiesterase (PDE) III inhibitor, blocked the effect of CO on I(Ba). Similar effects were seen in freshly dissociated human intestinal smooth muscle cells. The data suggest that exogenous CO can activate native and heterologously expressed intestinal L-type Ca(2+) channels through a pathway that involves activation of NOS, increased NO, and cGMP levels, but not PKG. Rather, the pathway appears to involve PKA, partly by reducing cAMP breakdown through inhibition of PDE III. CO-induced NO production may explain the apparent discrepancy between the low affinity of guanylyl cyclase for CO and the robust cGMP production evoked by CO.
Enteric NeuroScience Program, Mayo Clinic and Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
Inja
I
Lim
Simon J
SJ
Gibbons
Gregory L
GL
Lyford
Steven M
SM
Miller
Peter R
PR
Strege
Michael G
MG
Sarr
Suvro
S
Chatterjee
Joseph H
JH
Szurszewski
Vijay H
VH
Shah
Gianrico
G
Farrugia
D020746:Calcium Channels, L-Type / Q000187:drug effects* / Q000502:physiology*; D002248:Carbon Monoxide / Q000494:pharmacology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000494:pharmacology; D004068:Digestive System Physiological Phenomena; D005769:Gastrointestinal Motility; D006801:Humans; D007583:Jejunum / Q000166:cytology / Q000502:physiology; D009130:Muscle, Smooth / Q000502:physiology*; D009569:Nitric Oxide / Q000494:pharmacology*; D019001:Nitric Oxide Synthase / Q000494:pharmacology; D018408:Patch-Clamp Techniques
D020746:Calcium Channels, L-Type; D009569:Nitric Oxide; D002248:Carbon Monoxide; D019001:Nitric Oxide Synthase; D017868:Cyclic AMP-Dependent Protein Kinases