Transient receptor potential vanilloid 4-dependent calcium influx and ATP release in mouse and rat gastric epithelia

World J Gastroenterol. 2016 Jun 28;22(24):5512-9. doi: 10.3748/wjg.v22.i24.5512.

Abstract

Aim: To explore the expression of transient receptor potential vanilloid 4 (TRPV4) and its physiological meaning in mouse and rat gastric epithelia.

Methods: RT-PCR and immunochemistry were used to detect TRPV4 mRNA and protein expression in mouse stomach and a rat normal gastric epithelial cell line (RGE1-01), while Ca(2+)-imaging and electrophysiology were used to evaluate TRPV4 channel activity. ATP release was measured by a luciferin-luciferase assay. Gastric emptying was also compared between WT and TRPV4 knockout mice.

Results: TRPV4 mRNA and protein were detected in mouse tissues and RGE1-01 cells. A TRPV4-specific agonist (GSK1016790A) increased intracellular Ca(2+) concentrations and/or evoked TRPV4-like current activities in WT mouse gastric epithelial cells and RGE1-01 cells, but not TRPV4KO cells. GSK1016790A or mechanical stimuli induced ATP release from RGE1-01 cells while TRPV4 knockout mice displayed delayed gastric emptying in vivo.

Conclusion: TRPV4 is expressed in mouse and rat gastric epithelium and contributes to ATP release and gastric emptying.

Keywords: ATP; Gastric emptying; Stomach; Transient receptor potential vanilloid 4.

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Animals
  • Calcium / metabolism*
  • Cell Line
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism*
  • Gastric Emptying
  • Gastric Mucosa / metabolism*
  • Immunohistochemistry
  • Leucine / analogs & derivatives
  • Leucine / pharmacology
  • Mice
  • Mice, Knockout
  • Patch-Clamp Techniques
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sulfonamides / pharmacology
  • TRPV Cation Channels / agonists
  • TRPV Cation Channels / genetics*
  • TRPV Cation Channels / metabolism

Substances

  • N-(1-((4-(2-(((2,4-dichlorophenyl)sulfonyl)amino)-3-hydroxypropanoyl)-1-piperazinyl)carbonyl)-3-methylbutyl)-1-benzothiophene-2-carboxamide
  • Sulfonamides
  • TRPV Cation Channels
  • Trpv4 protein, mouse
  • Trpv4 protein, rat
  • Adenosine Triphosphate
  • Leucine
  • Calcium