Isoform-selective physical coupling of TRPC3 channels to IP3 receptors in smooth muscle cells regulates arterial contractility

Adebowale Adebiyi, Guiling Zhao, Damodaran Narayanan, Candice M. Thomas-Gatewood, John P. Bannister, Jonathan Jaggar

Research output: Contribution to journalArticle

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Abstract

RATIONALE: Inositol 1,4,5-trisphosphate (IP3)-induced vasoconstriction can occur independently of intracellular Ca release and via IP3 receptor (IP3R) and canonical transient receptor potential (TRPC) channel activation, but functional signaling mechanisms mediating this effect are unclear. OBJECTIVES: Study mechanisms by which IP 3Rs stimulate TRPC channels in myocytes of resistance-size cerebral arteries. METHODS AND RESULTS: Immunofluorescence resonance energy transfer (immuno-FRET) microscopy using isoform-selective antibodies indicated that endogenous type 1 IP3Rs (IP3R1) are in close spatial proximity to TRPC3, but distant from TRPC6 or TRPM4 channels in arterial myocytes. Endothelin-1 (ET-1), a phospholipase C-coupled receptor agonist, elevated immuno-FRET between IP3R1 and TRPC3, but not between IP3R1 and TRPC6 or TRPM4. TRPC3, but not TRPC6, coimmunoprecipitated with IP3R1. TRPC3 and TRPC6 antibodies selectively inhibited recombinant channels, but only the TRPC3 antibody blocked IP3-induced nonselective cation current (ICat) in myocytes. TRPC3 knockdown attenuated immuno-FRET between IP3R1 and TRPC3, IP3-induced ICat activation, and ET-1 and IP 3-induced vasoconstriction, whereas TRPC6 channel knockdown had no effect. ET-1 did not alter total or plasma membrane-localized TRPC3, as determined using surface biotinylation. RT-PCR demonstrated that C-terminal calmodulin and IP3R binding (CIRB) domains are present in myocyte TRPC3 and TRPC6 channels. A peptide corresponding to the IP3R N-terminal region that can interact with TRPC channels activated ICat. A TRPC3 CIRB domain peptide attenuated IP3-and ET-1-induced ICat activation and vasoconstriction. CONCLUSIONS: IP3 stimulates direct coupling between IP3R1 and membrane-resident TRPC3 channels in arterial myocytes, leading to ICat activation and vasoconstriction. Close spatial proximity between IP3R1 and TRPC3 establishes this isoform-selective functional interaction.

Original languageEnglish (US)
Pages (from-to)1603-1612
Number of pages10
JournalCirculation research
Volume106
Issue number10
DOIs
StatePublished - May 28 2010

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Inositol 1,4,5-Trisphosphate Receptors
Muscle Cells
Smooth Muscle Myocytes
Endothelin-1
Protein Isoforms
Vasoconstriction
Energy Transfer
Fluorescent Antibody Technique
Calmodulin
Antibodies
Biotinylation
Transient Receptor Potential Channels
Inositol 1,4,5-Trisphosphate
Cerebral Arteries
Type C Phospholipases
Cations
Microscopy
Cell Membrane
TRPC3 cation channel
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Isoform-selective physical coupling of TRPC3 channels to IP3 receptors in smooth muscle cells regulates arterial contractility. / Adebiyi, Adebowale; Zhao, Guiling; Narayanan, Damodaran; Thomas-Gatewood, Candice M.; Bannister, John P.; Jaggar, Jonathan.

In: Circulation research, Vol. 106, No. 10, 28.05.2010, p. 1603-1612.

Research output: Contribution to journalArticle

Adebiyi, Adebowale ; Zhao, Guiling ; Narayanan, Damodaran ; Thomas-Gatewood, Candice M. ; Bannister, John P. ; Jaggar, Jonathan. / Isoform-selective physical coupling of TRPC3 channels to IP3 receptors in smooth muscle cells regulates arterial contractility. In: Circulation research. 2010 ; Vol. 106, No. 10. pp. 1603-1612.
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AU - Adebiyi, Adebowale

AU - Zhao, Guiling

AU - Narayanan, Damodaran

AU - Thomas-Gatewood, Candice M.

AU - Bannister, John P.

AU - Jaggar, Jonathan

PY - 2010/5/28

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N2 - RATIONALE: Inositol 1,4,5-trisphosphate (IP3)-induced vasoconstriction can occur independently of intracellular Ca release and via IP3 receptor (IP3R) and canonical transient receptor potential (TRPC) channel activation, but functional signaling mechanisms mediating this effect are unclear. OBJECTIVES: Study mechanisms by which IP 3Rs stimulate TRPC channels in myocytes of resistance-size cerebral arteries. METHODS AND RESULTS: Immunofluorescence resonance energy transfer (immuno-FRET) microscopy using isoform-selective antibodies indicated that endogenous type 1 IP3Rs (IP3R1) are in close spatial proximity to TRPC3, but distant from TRPC6 or TRPM4 channels in arterial myocytes. Endothelin-1 (ET-1), a phospholipase C-coupled receptor agonist, elevated immuno-FRET between IP3R1 and TRPC3, but not between IP3R1 and TRPC6 or TRPM4. TRPC3, but not TRPC6, coimmunoprecipitated with IP3R1. TRPC3 and TRPC6 antibodies selectively inhibited recombinant channels, but only the TRPC3 antibody blocked IP3-induced nonselective cation current (ICat) in myocytes. TRPC3 knockdown attenuated immuno-FRET between IP3R1 and TRPC3, IP3-induced ICat activation, and ET-1 and IP 3-induced vasoconstriction, whereas TRPC6 channel knockdown had no effect. ET-1 did not alter total or plasma membrane-localized TRPC3, as determined using surface biotinylation. RT-PCR demonstrated that C-terminal calmodulin and IP3R binding (CIRB) domains are present in myocyte TRPC3 and TRPC6 channels. A peptide corresponding to the IP3R N-terminal region that can interact with TRPC channels activated ICat. A TRPC3 CIRB domain peptide attenuated IP3-and ET-1-induced ICat activation and vasoconstriction. CONCLUSIONS: IP3 stimulates direct coupling between IP3R1 and membrane-resident TRPC3 channels in arterial myocytes, leading to ICat activation and vasoconstriction. Close spatial proximity between IP3R1 and TRPC3 establishes this isoform-selective functional interaction.

AB - RATIONALE: Inositol 1,4,5-trisphosphate (IP3)-induced vasoconstriction can occur independently of intracellular Ca release and via IP3 receptor (IP3R) and canonical transient receptor potential (TRPC) channel activation, but functional signaling mechanisms mediating this effect are unclear. OBJECTIVES: Study mechanisms by which IP 3Rs stimulate TRPC channels in myocytes of resistance-size cerebral arteries. METHODS AND RESULTS: Immunofluorescence resonance energy transfer (immuno-FRET) microscopy using isoform-selective antibodies indicated that endogenous type 1 IP3Rs (IP3R1) are in close spatial proximity to TRPC3, but distant from TRPC6 or TRPM4 channels in arterial myocytes. Endothelin-1 (ET-1), a phospholipase C-coupled receptor agonist, elevated immuno-FRET between IP3R1 and TRPC3, but not between IP3R1 and TRPC6 or TRPM4. TRPC3, but not TRPC6, coimmunoprecipitated with IP3R1. TRPC3 and TRPC6 antibodies selectively inhibited recombinant channels, but only the TRPC3 antibody blocked IP3-induced nonselective cation current (ICat) in myocytes. TRPC3 knockdown attenuated immuno-FRET between IP3R1 and TRPC3, IP3-induced ICat activation, and ET-1 and IP 3-induced vasoconstriction, whereas TRPC6 channel knockdown had no effect. ET-1 did not alter total or plasma membrane-localized TRPC3, as determined using surface biotinylation. RT-PCR demonstrated that C-terminal calmodulin and IP3R binding (CIRB) domains are present in myocyte TRPC3 and TRPC6 channels. A peptide corresponding to the IP3R N-terminal region that can interact with TRPC channels activated ICat. A TRPC3 CIRB domain peptide attenuated IP3-and ET-1-induced ICat activation and vasoconstriction. CONCLUSIONS: IP3 stimulates direct coupling between IP3R1 and membrane-resident TRPC3 channels in arterial myocytes, leading to ICat activation and vasoconstriction. Close spatial proximity between IP3R1 and TRPC3 establishes this isoform-selective functional interaction.

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