. TRPC5 as a candidate for the nonselective cation channel activated by muscarinic stimulation in murine stomach. Am J Physiol Gastrointest Liver Physiol 284: G604-G616, 2003; 10.1152/ajpgi.00069.2002.-We investigated which transient receptor potential (TRP) channel is responsible for the nonselective cation channel (NSCC) activated by carbachol (CCh) in murine stomach with RT-PCR and the electrophysiological method. All seven types of TRP mRNA were detected in murine stomach with RT-PCR. When each TRP channel was expressed, the current-voltage relationship of mTRP5 was most similar to that recorded in murine gastric myocytes. mTRP5 showed a conductance order of Cs ϩ Ͼ K ϩ Ͼ Na ϩ , similar to that in the murine stomach. With 0.2 mM GTP␥S in the pipette solution, the current was activated transiently in both NSCC in the murine stomach and the expressed mTRP5. Both NSCC activated by CCh in murine stomach and mTRP5 were inhibited by intracellularly applied anti-G q/11 antibody, PLC inhibitor U-73122, IICR inhibitor 2-aminoethoxydiphenylborate, and nonspecific cation channel blockers La 3ϩ and flufenamate. There were two other unique properties. Both the native NSCC and mTRP5 were activated by 1-oleoyl-2-acetyl-sn-glycerol. Without the activation of NSCC by CCh, the NSCC in murine stomach was constitutively active like mTRP5. From the above results, we suggest that mTRP5 might be a candidate for the NSCC activated by ACh or CCh in murine stomach. transient receptor potential protein

No abstract

Interstitial cells of Cajal (ICCs) are a fundamental component of the pacemaker apparatus of the gastrointestinal (GI) tract. They have special properties that make them unique in their ability to generate and propagate slow waves in gastrointestinal smooth muscle. The pacemaker current that generates slow waves is initially due to a voltageindependent, Ca 2+ -inhibited, non-selective cationic conductance in ICC. The classical transient receptor potential (TRPC) channel 4 was suggested as a molecular candidate for the nonselective cation channel (NSCC) responsible for the pacemaker activity. We have shown that TRPC4-/-mice display normal slow waves and suggest that TRPC4 might be an essential component of the NSCC activated by muscarinic stimulation. Finally, we suggest that TRPM7 is the molecular candidate for the NSCC responsible for pacemaker activity in ICCs on the basis of electrophysiological, molecular biological, and immunohistochemical experiments.

In this study, we investigated which subtype of GTP-binding protein (G protein) is related to muscarinic activation of nonselective cation (NSC) channels in gastric smooth muscle. Inward cationic current was activated by the application of 50 microM carbachol (ICCh) at a holding potential of -60 mV with the same CsCl-rich solution in both pipette and bath. The same cationic current as ICCh was slowly activated by the dialysis of guanosine 5'-O-(3-thiotriphosphate) (GTP[gamma-S]) through the pipette. Since it is known that pertussis toxin pretreatment can block ICCh, antibodies (Abs) against Galpha,i (anti-Galpha,i) or Galpha,o (anti-Galpha,o) were tested. Activation of ICCh was blocked by the addition of anti-Galpha,o. However, anti-Galpha,i Abs had no significant effect on ICCh. The expression of Galpha,o in guinea-pig gastric smooth muscle was confirmed by Western immunoblot analysis. These results suggest that Go-type protein may mediate signals from the muscarinic receptor to NSC channel in guinea-pig gastric myocytes.