CaV1.2 channel N-terminal splice variants modulate functional surface expression in resistance size artery smooth muscle cells

John P. Bannister, Candice M. Thomas-Gatewood, Zachary P. Neeb, Adebowale Adebiyi, Xiaoyang Cheng, Jonathan Jaggar

Research output: Contribution to journalArticle

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Abstract

Voltage-dependent Ca2+ (CaV1.2) channels are the primary Ca2+ influx pathway in arterial smooth muscle cells and are essential for contractility regulation by a variety of stimuli, including intravascular pressure. Arterial smooth muscle cell CaV1.2 mRNA is alternatively spliced at exon 1 (e1), generating e1b or e1c variants, with e1c exhibiting relatively smooth muscle-specific expression in the cardiovascular system. Here, we examined physiological functions of CaV1.2e1 variants and tested the hypothesis that targeting CaV1.2e1 modulates resistance size cerebral artery contractility. Custom antibodies that selectively recognize CaV1.2 channel proteins containing sequences encoded by either e1b (CaV1.2e1b) or e1c (CaV1.2e1c) both detected CaV1.2 in rat and human cerebral arteries. shRNA targeting e1b or e1c reduced expression of that CaV1.2 variant, induced compensatory up-regulation of the other variant, decreased total Ca V1.2, and reduced intravascular pressure-and depolarization-induced vasoconstriction. CaV1.2e1b and CaV1.2e1c knockdown reduced whole cell CaV1.2 currents, with CaV1.2e1c knockdown most effectively reducing total CaV1.2 and inducing the largest vasodilation. Knockdown of α2Δ-1, a Ca V1.2 auxiliary subunit, reduced surface expression of both Ca V1.2e1 variants, inhibiting CaV1.2e1c more than Ca V1.2e1b. e1b or e1c overexpression reduced CaV1.2 surface expression and whole cell currents, leading to vasodilation, with e1c overexpression inducing the largest effect. In summary, data indicate that arterial smooth muscle cells express CaV1.2 channels containing e1b or e1c-encoded N termini that contribute to CaV1.2 surface expression, α2S-1 preferentially traffics the Ca V1.2e1c variant to the plasma membrane, and targeting of Ca V1.2e1 message or the CaV1.2 channel proximal N terminus induces vasodilation.

Original languageEnglish (US)
Pages (from-to)15058-15066
Number of pages9
JournalJournal of Biological Chemistry
Volume286
Issue number17
DOIs
StatePublished - Apr 29 2011

Fingerprint

Vasodilation
Smooth Muscle Myocytes
Muscle
Cerebral Arteries
Arteries
Cells
Pressure
Cardiovascular System
Vasoconstriction
Cardiovascular system
Small Interfering RNA
Smooth Muscle
Exons
Depolarization
Up-Regulation
Cell membranes
Cell Membrane
Rats
Messenger RNA
Antibodies

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

CaV1.2 channel N-terminal splice variants modulate functional surface expression in resistance size artery smooth muscle cells. / Bannister, John P.; Thomas-Gatewood, Candice M.; Neeb, Zachary P.; Adebiyi, Adebowale; Cheng, Xiaoyang; Jaggar, Jonathan.

In: Journal of Biological Chemistry, Vol. 286, No. 17, 29.04.2011, p. 15058-15066.

Research output: Contribution to journalArticle

Bannister, John P. ; Thomas-Gatewood, Candice M. ; Neeb, Zachary P. ; Adebiyi, Adebowale ; Cheng, Xiaoyang ; Jaggar, Jonathan. / CaV1.2 channel N-terminal splice variants modulate functional surface expression in resistance size artery smooth muscle cells. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 17. pp. 15058-15066.
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AU - Adebiyi, Adebowale

AU - Cheng, Xiaoyang

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