Hydrogen sulfide is a partially redox-independent activator of the human jejunum Na+ channel, NAv1.5

Peter R. Strege, Cheryl E. Bernard, Robert E. Kraichely, Amelia Mazzone, Lei Sha, Arthur Beyder, Simon J. Gibbons, David R Linden, Michael L. Kendrick, Michael G. Sarr, Joseph H. Szurszewski, Gianrico Farrugia

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Hydrogen sulfide (H2S) is produced endogenously by L-cysteine metabolism. H2S modulates several ion channels with an unclear mechanism of action. A possible mechanism is through reduction-oxidation reactions attributable to the redox potential of the sulfur moiety. The aims of this study were to determine the effects of the H2S donor NaHS on Nav1.5, a voltage-dependent sodium channel expressed in the gastrointestinal tract in human jejunum smooth muscle cells and interstitial cells of Cajal, and to elucidate whether H2S acts on Nav1.5 by redox reactions. Whole cell Na+ currents were recorded in freshly dissociated human jejunum circular myocytes and Nav1.5-transfected human embryonic kidney-293 cells. RT-PCR amplified mRNA for H2S enzymes cystathionine β-synthase and cystathionine γ-lyase from the human jejunum. NaHS increased native Na+ peak currents and shifted the half-point (V1/2) of steady-state activation and inactivation by +21 ± 2 mV and +15 ± 3 mV, respectively. Similar effects were seen on the heterologously expressed NaV1.5 α subunit with EC50s in the 10-4 to 10-3 M range. The reducing agent dithiothreitol (DTT) mimicked in part the effects of NaHS by increasing peak current and positively shifting steady-state activation. DTT together with NaHS had an additive effect on steady-state activation but not on peak current, suggesting that the latter may be altered via reduction. Pretreatment with the Hg2+-conjugated oxidizer thimerosal or the alkylating agent N-ethylmaleimide inhibited or decreased NaHS induction of Nav1.5 peak current. These studies show that H2S activates the gastrointestinal Na+ channel, and the mechanism of action of H2S is partially redox independent.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume300
Issue number6
DOIs
StatePublished - Jun 2011

Fingerprint

Hydrogen Sulfide
Jejunum
Oxidation-Reduction
Cystathionine
Dithiothreitol
Interstitial Cells of Cajal
Thimerosal
Ethylmaleimide
Lyases
Sodium Channels
Alkylating Agents
Reducing Agents
Ion Channels
Sulfur
Muscle Cells
Smooth Muscle Myocytes
Cysteine
Gastrointestinal Tract
sodium bisulfide
Kidney

Keywords

  • Dithiothreitol
  • Intestine
  • Patch clamp
  • Smooth muscle
  • Thimerosal

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology (medical)
  • Physiology
  • Hepatology

Cite this

Hydrogen sulfide is a partially redox-independent activator of the human jejunum Na+ channel, NAv1.5. / Strege, Peter R.; Bernard, Cheryl E.; Kraichely, Robert E.; Mazzone, Amelia; Sha, Lei; Beyder, Arthur; Gibbons, Simon J.; Linden, David R; Kendrick, Michael L.; Sarr, Michael G.; Szurszewski, Joseph H.; Farrugia, Gianrico.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 300, No. 6, 06.2011.

Research output: Contribution to journalArticle

Strege, Peter R. ; Bernard, Cheryl E. ; Kraichely, Robert E. ; Mazzone, Amelia ; Sha, Lei ; Beyder, Arthur ; Gibbons, Simon J. ; Linden, David R ; Kendrick, Michael L. ; Sarr, Michael G. ; Szurszewski, Joseph H. ; Farrugia, Gianrico. / Hydrogen sulfide is a partially redox-independent activator of the human jejunum Na+ channel, NAv1.5. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2011 ; Vol. 300, No. 6.
@article{821f1ff2dad6401eb37e6ba2e30a27e3,
title = "Hydrogen sulfide is a partially redox-independent activator of the human jejunum Na+ channel, NAv1.5",
abstract = "Hydrogen sulfide (H2S) is produced endogenously by L-cysteine metabolism. H2S modulates several ion channels with an unclear mechanism of action. A possible mechanism is through reduction-oxidation reactions attributable to the redox potential of the sulfur moiety. The aims of this study were to determine the effects of the H2S donor NaHS on Nav1.5, a voltage-dependent sodium channel expressed in the gastrointestinal tract in human jejunum smooth muscle cells and interstitial cells of Cajal, and to elucidate whether H2S acts on Nav1.5 by redox reactions. Whole cell Na+ currents were recorded in freshly dissociated human jejunum circular myocytes and Nav1.5-transfected human embryonic kidney-293 cells. RT-PCR amplified mRNA for H2S enzymes cystathionine β-synthase and cystathionine γ-lyase from the human jejunum. NaHS increased native Na+ peak currents and shifted the half-point (V1/2) of steady-state activation and inactivation by +21 ± 2 mV and +15 ± 3 mV, respectively. Similar effects were seen on the heterologously expressed NaV1.5 α subunit with EC50s in the 10-4 to 10-3 M range. The reducing agent dithiothreitol (DTT) mimicked in part the effects of NaHS by increasing peak current and positively shifting steady-state activation. DTT together with NaHS had an additive effect on steady-state activation but not on peak current, suggesting that the latter may be altered via reduction. Pretreatment with the Hg2+-conjugated oxidizer thimerosal or the alkylating agent N-ethylmaleimide inhibited or decreased NaHS induction of Nav1.5 peak current. These studies show that H2S activates the gastrointestinal Na+ channel, and the mechanism of action of H2S is partially redox independent.",
keywords = "Dithiothreitol, Intestine, Patch clamp, Smooth muscle, Thimerosal",
author = "Strege, {Peter R.} and Bernard, {Cheryl E.} and Kraichely, {Robert E.} and Amelia Mazzone and Lei Sha and Arthur Beyder and Gibbons, {Simon J.} and Linden, {David R} and Kendrick, {Michael L.} and Sarr, {Michael G.} and Szurszewski, {Joseph H.} and Gianrico Farrugia",
year = "2011",
month = "6",
doi = "10.1152/ajpgi.00556.2010",
language = "English (US)",
volume = "300",
journal = "American Journal of Physiology - Renal Fluid and Electrolyte Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "6",

}

TY - JOUR

T1 - Hydrogen sulfide is a partially redox-independent activator of the human jejunum Na+ channel, NAv1.5

AU - Strege, Peter R.

AU - Bernard, Cheryl E.

AU - Kraichely, Robert E.

AU - Mazzone, Amelia

AU - Sha, Lei

AU - Beyder, Arthur

AU - Gibbons, Simon J.

AU - Linden, David R

AU - Kendrick, Michael L.

AU - Sarr, Michael G.

AU - Szurszewski, Joseph H.

AU - Farrugia, Gianrico

PY - 2011/6

Y1 - 2011/6

N2 - Hydrogen sulfide (H2S) is produced endogenously by L-cysteine metabolism. H2S modulates several ion channels with an unclear mechanism of action. A possible mechanism is through reduction-oxidation reactions attributable to the redox potential of the sulfur moiety. The aims of this study were to determine the effects of the H2S donor NaHS on Nav1.5, a voltage-dependent sodium channel expressed in the gastrointestinal tract in human jejunum smooth muscle cells and interstitial cells of Cajal, and to elucidate whether H2S acts on Nav1.5 by redox reactions. Whole cell Na+ currents were recorded in freshly dissociated human jejunum circular myocytes and Nav1.5-transfected human embryonic kidney-293 cells. RT-PCR amplified mRNA for H2S enzymes cystathionine β-synthase and cystathionine γ-lyase from the human jejunum. NaHS increased native Na+ peak currents and shifted the half-point (V1/2) of steady-state activation and inactivation by +21 ± 2 mV and +15 ± 3 mV, respectively. Similar effects were seen on the heterologously expressed NaV1.5 α subunit with EC50s in the 10-4 to 10-3 M range. The reducing agent dithiothreitol (DTT) mimicked in part the effects of NaHS by increasing peak current and positively shifting steady-state activation. DTT together with NaHS had an additive effect on steady-state activation but not on peak current, suggesting that the latter may be altered via reduction. Pretreatment with the Hg2+-conjugated oxidizer thimerosal or the alkylating agent N-ethylmaleimide inhibited or decreased NaHS induction of Nav1.5 peak current. These studies show that H2S activates the gastrointestinal Na+ channel, and the mechanism of action of H2S is partially redox independent.

AB - Hydrogen sulfide (H2S) is produced endogenously by L-cysteine metabolism. H2S modulates several ion channels with an unclear mechanism of action. A possible mechanism is through reduction-oxidation reactions attributable to the redox potential of the sulfur moiety. The aims of this study were to determine the effects of the H2S donor NaHS on Nav1.5, a voltage-dependent sodium channel expressed in the gastrointestinal tract in human jejunum smooth muscle cells and interstitial cells of Cajal, and to elucidate whether H2S acts on Nav1.5 by redox reactions. Whole cell Na+ currents were recorded in freshly dissociated human jejunum circular myocytes and Nav1.5-transfected human embryonic kidney-293 cells. RT-PCR amplified mRNA for H2S enzymes cystathionine β-synthase and cystathionine γ-lyase from the human jejunum. NaHS increased native Na+ peak currents and shifted the half-point (V1/2) of steady-state activation and inactivation by +21 ± 2 mV and +15 ± 3 mV, respectively. Similar effects were seen on the heterologously expressed NaV1.5 α subunit with EC50s in the 10-4 to 10-3 M range. The reducing agent dithiothreitol (DTT) mimicked in part the effects of NaHS by increasing peak current and positively shifting steady-state activation. DTT together with NaHS had an additive effect on steady-state activation but not on peak current, suggesting that the latter may be altered via reduction. Pretreatment with the Hg2+-conjugated oxidizer thimerosal or the alkylating agent N-ethylmaleimide inhibited or decreased NaHS induction of Nav1.5 peak current. These studies show that H2S activates the gastrointestinal Na+ channel, and the mechanism of action of H2S is partially redox independent.

KW - Dithiothreitol

KW - Intestine

KW - Patch clamp

KW - Smooth muscle

KW - Thimerosal

UR - http://www.scopus.com/inward/record.url?scp=79957929115&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79957929115&partnerID=8YFLogxK

U2 - 10.1152/ajpgi.00556.2010

DO - 10.1152/ajpgi.00556.2010

M3 - Article

C2 - 21393430

AN - SCOPUS:79957929115

VL - 300

JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

SN - 1931-857X

IS - 6

ER -