Chonnam Med J.  2011 Aug;47(2):72-79. 10.4068/cmj.2011.47.2.72.

Interplay of Hydrogen Sulfide and Nitric Oxide on the Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine

Affiliations
  • 1Department of Physiology, College of Medicine, Chosun University, Gwangju, Korea. jyjun@chosun.ac.kr

Abstract

We studied whether nitric oxide (NO) and hydrogen sulfide (H2S) have an interaction on the pacemaker activities of interstitial cells of Cajal (ICC) from the mouse small intestine. The actions of NO and H2S on pacemaker activities were investigated by using the whole-cell patch-clamp technique and intracellular Ca2+ analysis at 30degrees C in cultured mouse ICC. Exogenously applied (+/-)-S-nitroso-N-acetylpenicillamine (SNAP), an NO donor, or sodium hydrogen sulfide (NaHS), a donor of H2S, showed no influence on pacemaker activity (potentials and currents) in ICC at low concentrations (10 microM SNAP and 100 microM NaHS), but SNAP or NaHS completely inhibited pacemaker amplitude and pacemaker frequency with increases in the resting currents in the outward direction at high concentrations (SNAP 100 microM and NaHS 1 mM). Co-treatment with 10 microM SNAP plus 100 microM NaHS also inhibited pacemaker amplitude and pacemaker frequency with increases in the resting currents in the outward direction. ODQ, a guanylate cyclase inhibitor, or glibenclamide, an ATP-sensitive K+ channel inhibitor, blocked the SNAP+NaHS-induced inhibition of pacemaker currents in ICC. Also, we found that SNAP+NaHS inhibited the spontaneous intracellular Ca2+ ([Ca2+]i) oscillations in cultured ICC. In conclusion, this study describes the enhanced inhibitory effects of NO plus H2S on ICC in the mouse small intestine. NO+H2S inhibited the pacemaker activity of ICC by modulating intracellular Ca2+. These results may be evidence of a physiological interaction of NO and H2S in ICC for modulating gastrointestinal motility.

Keyword

Interstitial cells of Cajal (ICC); Intestinal motility; Pacemaker currents; Nitric Oxide; Hydrogen sulfide

MeSH Terms

Animals
Gastrointestinal Motility
Glyburide
Guanylate Cyclase
Humans
Hydrogen
Hydrogen Sulfide
Interstitial Cells of Cajal
Intestine, Small
Mice
Nitric Oxide
Patch-Clamp Techniques
Sodium
Sulfides
Tissue Donors
Glyburide
Guanylate Cyclase
Hydrogen
Hydrogen Sulfide
Nitric Oxide
Sodium
Sulfides

Figure

  • FIG. 1 Effects of H2S+NO on pacemaker potentials recorded in ICC. (A) Pacemaker potentials of ICC exposed to SNAP (10 µM) in the current clamping mode (I=0). (B) Pacemaker potentials of ICC exposed to NaHS (100 µM) in the current clamping mode (I=0). (C) Pacemaker potentials of ICC exposed to SNAP (10 µM)+NaHS (100 µM) in the current clamping mode (I=0). The dotted lines indicate the control resting membrane potentials. Responses to SNAP+NaHS are summarized in (D, E). Bars represent the means±SE. *Asterisks indicate significantly different from controls (p<0.05). Con: control, RMP: resting membrane potentials.

  • FIG. 2 Effects of H2S+NO on pacemaker currents recorded in cultured ICC. (A) Pacemaker currents of ICC exposed to SNAP (100 µM) at a holding potential of -70 mV. (B) Pacemaker currents of ICC exposed to NaHS (1 mM) at a holding potential of -70 mV. (C) Pacemaker currents of ICC exposed to SNAP (10 µM) at a holding potential of -70 mV. (D) Pacemaker currents of ICC exposed to NaHS (100 µM) at a holding potential of -70 mV. (E) Pacemaker potentials of ICC exposed to SNAP (10 µM)+NaHS (100 µM) at a holding potential of -70 mV. The dotted lines indicate the control resting current levels. (F-H) summarize the effects of SNAP+NaHS on pacemaker currents in ICC. Bars represent the means±SE. *Asterisks indicate significantly different from single treatment of SNAP (10 µM) or NaHS (100 µM) (p<0.05). Con: control.

  • FIG. 3 Effects of ODQ or glibenclamide on NO+H2S-induced effects in cultured ICC. (A) Pacemaker currents of ICC exposed to SNAP (10 µM)+NaHS (100 µM) in the presence of ODQ (10 µM). (B) Pacemaker currents of ICC exposed to SNAP (10 µM)+NaHS (100 µM) in the presence of glibenclamide (10 µM). Responses to SNAP+NaHS in the presence of ODQ or glibenclamide are summarized in (C-E). Bars represent the mean values±SE. The dotted lines indicate the zero current levels. *Asterisks indicate significantly different from SNAP+NaHS (p<0.05). Con: control, GBC: glibenclamide.

  • FIG. 4 Effects of NO+H2S on [Ca2+]i oscillation in cultured ICC. (A) Basal and peak point of the ICC image in the presence of SNAP (10 µM). (B) The sequential fluorescence intensity change plotted in (A) is shown in red. (C) Basal and peak point of the ICC image in the presence of NaHS (100 µM). (D) The sequential fluorescence intensity change plotted in (C) is shown in red. (E) Basal and peak point of the ICC image in the control condition. (F) Basal and peak point of the ICC image in the presence of SNAP (10 µM)+NaHS (100 µM). (G) The sequential fluorescence intensity change plotted in (E, F) is shown in red. The interval of the representative frame was 1 s and the exposure time of each frame was 500 ms.


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