Smooth muscle cell α2δ-1 subunits are essential for vasoregulation by CaV1.2 channels

John P. Bannister, Adebowale Adebiyi, Guiling Zhao, Damodaran Narayanan, Candice M. Thomas, Jessie Y. Feng, Jonathan Jaggar

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Rationale: Voltage-dependent L-type (CaV1.2) Ca2+ channels are a heteromeric complex formed from pore-forming α1 and auxiliary α2δ and β subunits. CaV1.2 channels are the principal Ca2+ influx pathway in arterial myocytes and regulate multiple physiological functions, including contraction. The macromolecular composition of arterial myocyte CaV1.2 channels remains poorly understood, with no studies having examined the molecular identity or physiological functions of α2δ subunits. Objective: We investigated the functional significance of α2δ subunits in myocytes of resistance-size (100 to 200 μm diameter) cerebral arteries. Methods and Results: α2δ-1 was the only α2δ isoform expressed in cerebral artery myocytes. Pregabalin, an α2δ-1/-2 ligand, and an α2δ-1 antibody, inhibited CaV1.2 currents in isolated myocytes. Acute pregabalin application reversibly dilated pressurized arteries. Using a novel application of surface biotinylation, data indicated that >95% of CaV1.2 α1 and α2δ-1 subunits were present in the arterial myocyte plasma membrane. α2δ-1 knockdown using short hairpin RNA reduced plasma membrane-localized Ca V1.2 α1 subunits, caused a corresponding elevation in cytosolic CaV1.2 α1 subunits, decreased intracellular Ca2+ concentration, inhibited pressure-induced vasoconstriction (myogenic tone), and attenuated pregabalin-induced vasodilation. Prolonged (24-hour) pregabalin exposure did not alter total α2δ-1 or CaV1.2 α1 proteins but decreased plasma membrane expression of each subunit, which reduced myogenic tone. Conclusions: α2δ-1 is essential for plasma membrane expression of arterial myocyte Ca V1.2 α1 subunits. α2δ-1 targeting can block CaV1.2 channels directly and inhibit surface expression of CaV1.2 α1 subunits, leading to vasodilation. These data identify α2δ-1 as a novel molecular target in arterial myocytes, the manipulation of which regulates contractility.

Original languageEnglish (US)
Pages (from-to)948-955
Number of pages8
JournalCirculation research
Volume105
Issue number10
DOIs
StatePublished - Jan 1 2009

Fingerprint

Muscle Cells
Smooth Muscle Myocytes
Cell Membrane
Cerebral Arteries
Vasodilation
Biotinylation
Vasoconstriction
Small Interfering RNA
Protein Isoforms
Arteries
Ligands
Pressure
Pregabalin
Antibodies
Proteins

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Smooth muscle cell α2δ-1 subunits are essential for vasoregulation by CaV1.2 channels. / Bannister, John P.; Adebiyi, Adebowale; Zhao, Guiling; Narayanan, Damodaran; Thomas, Candice M.; Feng, Jessie Y.; Jaggar, Jonathan.

In: Circulation research, Vol. 105, No. 10, 01.01.2009, p. 948-955.

Research output: Contribution to journalArticle

Bannister, John P. ; Adebiyi, Adebowale ; Zhao, Guiling ; Narayanan, Damodaran ; Thomas, Candice M. ; Feng, Jessie Y. ; Jaggar, Jonathan. / Smooth muscle cell α2δ-1 subunits are essential for vasoregulation by CaV1.2 channels. In: Circulation research. 2009 ; Vol. 105, No. 10. pp. 948-955.
@article{03f32202ab0a426eb14aa8e76549f242,
title = "Smooth muscle cell α2δ-1 subunits are essential for vasoregulation by CaV1.2 channels",
abstract = "Rationale: Voltage-dependent L-type (CaV1.2) Ca2+ channels are a heteromeric complex formed from pore-forming α1 and auxiliary α2δ and β subunits. CaV1.2 channels are the principal Ca2+ influx pathway in arterial myocytes and regulate multiple physiological functions, including contraction. The macromolecular composition of arterial myocyte CaV1.2 channels remains poorly understood, with no studies having examined the molecular identity or physiological functions of α2δ subunits. Objective: We investigated the functional significance of α2δ subunits in myocytes of resistance-size (100 to 200 μm diameter) cerebral arteries. Methods and Results: α2δ-1 was the only α2δ isoform expressed in cerebral artery myocytes. Pregabalin, an α2δ-1/-2 ligand, and an α2δ-1 antibody, inhibited CaV1.2 currents in isolated myocytes. Acute pregabalin application reversibly dilated pressurized arteries. Using a novel application of surface biotinylation, data indicated that >95{\%} of CaV1.2 α1 and α2δ-1 subunits were present in the arterial myocyte plasma membrane. α2δ-1 knockdown using short hairpin RNA reduced plasma membrane-localized Ca V1.2 α1 subunits, caused a corresponding elevation in cytosolic CaV1.2 α1 subunits, decreased intracellular Ca2+ concentration, inhibited pressure-induced vasoconstriction (myogenic tone), and attenuated pregabalin-induced vasodilation. Prolonged (24-hour) pregabalin exposure did not alter total α2δ-1 or CaV1.2 α1 proteins but decreased plasma membrane expression of each subunit, which reduced myogenic tone. Conclusions: α2δ-1 is essential for plasma membrane expression of arterial myocyte Ca V1.2 α1 subunits. α2δ-1 targeting can block CaV1.2 channels directly and inhibit surface expression of CaV1.2 α1 subunits, leading to vasodilation. These data identify α2δ-1 as a novel molecular target in arterial myocytes, the manipulation of which regulates contractility.",
author = "Bannister, {John P.} and Adebowale Adebiyi and Guiling Zhao and Damodaran Narayanan and Thomas, {Candice M.} and Feng, {Jessie Y.} and Jonathan Jaggar",
year = "2009",
month = "1",
day = "1",
doi = "10.1161/CIRCRESAHA.109.203620",
language = "English (US)",
volume = "105",
pages = "948--955",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "Lippincott Williams and Wilkins",
number = "10",

}

TY - JOUR

T1 - Smooth muscle cell α2δ-1 subunits are essential for vasoregulation by CaV1.2 channels

AU - Bannister, John P.

AU - Adebiyi, Adebowale

AU - Zhao, Guiling

AU - Narayanan, Damodaran

AU - Thomas, Candice M.

AU - Feng, Jessie Y.

AU - Jaggar, Jonathan

PY - 2009/1/1

Y1 - 2009/1/1

N2 - Rationale: Voltage-dependent L-type (CaV1.2) Ca2+ channels are a heteromeric complex formed from pore-forming α1 and auxiliary α2δ and β subunits. CaV1.2 channels are the principal Ca2+ influx pathway in arterial myocytes and regulate multiple physiological functions, including contraction. The macromolecular composition of arterial myocyte CaV1.2 channels remains poorly understood, with no studies having examined the molecular identity or physiological functions of α2δ subunits. Objective: We investigated the functional significance of α2δ subunits in myocytes of resistance-size (100 to 200 μm diameter) cerebral arteries. Methods and Results: α2δ-1 was the only α2δ isoform expressed in cerebral artery myocytes. Pregabalin, an α2δ-1/-2 ligand, and an α2δ-1 antibody, inhibited CaV1.2 currents in isolated myocytes. Acute pregabalin application reversibly dilated pressurized arteries. Using a novel application of surface biotinylation, data indicated that >95% of CaV1.2 α1 and α2δ-1 subunits were present in the arterial myocyte plasma membrane. α2δ-1 knockdown using short hairpin RNA reduced plasma membrane-localized Ca V1.2 α1 subunits, caused a corresponding elevation in cytosolic CaV1.2 α1 subunits, decreased intracellular Ca2+ concentration, inhibited pressure-induced vasoconstriction (myogenic tone), and attenuated pregabalin-induced vasodilation. Prolonged (24-hour) pregabalin exposure did not alter total α2δ-1 or CaV1.2 α1 proteins but decreased plasma membrane expression of each subunit, which reduced myogenic tone. Conclusions: α2δ-1 is essential for plasma membrane expression of arterial myocyte Ca V1.2 α1 subunits. α2δ-1 targeting can block CaV1.2 channels directly and inhibit surface expression of CaV1.2 α1 subunits, leading to vasodilation. These data identify α2δ-1 as a novel molecular target in arterial myocytes, the manipulation of which regulates contractility.

AB - Rationale: Voltage-dependent L-type (CaV1.2) Ca2+ channels are a heteromeric complex formed from pore-forming α1 and auxiliary α2δ and β subunits. CaV1.2 channels are the principal Ca2+ influx pathway in arterial myocytes and regulate multiple physiological functions, including contraction. The macromolecular composition of arterial myocyte CaV1.2 channels remains poorly understood, with no studies having examined the molecular identity or physiological functions of α2δ subunits. Objective: We investigated the functional significance of α2δ subunits in myocytes of resistance-size (100 to 200 μm diameter) cerebral arteries. Methods and Results: α2δ-1 was the only α2δ isoform expressed in cerebral artery myocytes. Pregabalin, an α2δ-1/-2 ligand, and an α2δ-1 antibody, inhibited CaV1.2 currents in isolated myocytes. Acute pregabalin application reversibly dilated pressurized arteries. Using a novel application of surface biotinylation, data indicated that >95% of CaV1.2 α1 and α2δ-1 subunits were present in the arterial myocyte plasma membrane. α2δ-1 knockdown using short hairpin RNA reduced plasma membrane-localized Ca V1.2 α1 subunits, caused a corresponding elevation in cytosolic CaV1.2 α1 subunits, decreased intracellular Ca2+ concentration, inhibited pressure-induced vasoconstriction (myogenic tone), and attenuated pregabalin-induced vasodilation. Prolonged (24-hour) pregabalin exposure did not alter total α2δ-1 or CaV1.2 α1 proteins but decreased plasma membrane expression of each subunit, which reduced myogenic tone. Conclusions: α2δ-1 is essential for plasma membrane expression of arterial myocyte Ca V1.2 α1 subunits. α2δ-1 targeting can block CaV1.2 channels directly and inhibit surface expression of CaV1.2 α1 subunits, leading to vasodilation. These data identify α2δ-1 as a novel molecular target in arterial myocytes, the manipulation of which regulates contractility.

UR - http://www.scopus.com/inward/record.url?scp=72449122873&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=72449122873&partnerID=8YFLogxK

U2 - 10.1161/CIRCRESAHA.109.203620

DO - 10.1161/CIRCRESAHA.109.203620

M3 - Article

VL - 105

SP - 948

EP - 955

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 10

ER -