Type 1 IP3 receptors activate BKC3 channels via local molecular coupling in arterial smooth muscle cells

Guiling Zhao, Zachary P. Neeb, Marie Dennis Leo, Judith Pachuau, Adebowale Adebiyi, Kunfu Ouyang, Ju Chen, Jonathan Jaggar

Research output: Contribution to journalArticle

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Abstract

Plasma membrane large-conductance Ca2+-activated K +(BKC3) channels and sarcoplasmic reticulum inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) are expressed in a wide variety of cell types, including arterial smooth muscle cells. Here, we studied BKCa channel regulation by IP3 and IP 3Rs in rat and mouse cerebral artery smooth muscle cells. IP 3 activated BKCa channels both in intact cells and in excised inside-out membrane patches. IP3 caused concentration- dependent BKCa channel activation with an apparent dissociation constant (Kd) of ∼4 μM at physiological voltage (-40 mV) and intracellular Ca2+ concentration ( [Ca2+]i; 10 μM). IP3 also caused a leftward-shift in BKCa channel apparent Ca2+ sensitivity and reduced the Kd for free [Ca2+i from ∼20 to 12 μM, but did not alter the slope or maximal Po. BAPTA, a fast Ca2+ buffer, or an elevation in extracellular Ca2+ concentration did not alter IPs-induced BKCa channel activation. Heparin, an IP3R inhibitor, and a monoclonal type 1 IP3R (IP3R1) antibody blocked IP3-induced BKCa channel activation. Adenophostin A, an IP3R agonist, also activated BKCa channels. IP 3 activated BKCa channels in inside-out patches from wild-type (IP3R1+/+) mouse arterial smooth muscle cells, but had no effect on BKCa channels of IP2R1-deficient (IP3R1-/-) mice. Immunofluorescence resonance energy transfer microscopy indicated that IP3R1 is located in close spatial proximity to BKCa α subunits. The IP3R1 monoclonal antibody coimmunoprecipitated IP3R1 and BKCa, channel α and β1 subunits from cerebral arteries. In summary, data indicate that IP3R1 activation elevates BKCa channel apparent Ca2+ sensitivity through local molecular coupling in arterial smooth muscle cells.

Original languageEnglish (US)
Pages (from-to)283-291
Number of pages9
JournalJournal of General Physiology
Volume136
Issue number3
DOIs
StatePublished - Sep 1 2010

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Inositol 1,4,5-Trisphosphate Receptors
Smooth Muscle Myocytes
Cerebral Arteries
Calcium-Activated Potassium Channels
Inositol 1,4,5-Trisphosphate
Energy Transfer
Sarcoplasmic Reticulum
Fluorescent Antibody Technique
Microscopy
Buffers
Monoclonal Antibodies
Cell Membrane
Membranes
Antibodies

All Science Journal Classification (ASJC) codes

  • Physiology

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Type 1 IP3 receptors activate BKC3 channels via local molecular coupling in arterial smooth muscle cells. / Zhao, Guiling; Neeb, Zachary P.; Leo, Marie Dennis; Pachuau, Judith; Adebiyi, Adebowale; Ouyang, Kunfu; Chen, Ju; Jaggar, Jonathan.

In: Journal of General Physiology, Vol. 136, No. 3, 01.09.2010, p. 283-291.

Research output: Contribution to journalArticle

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abstract = "Plasma membrane large-conductance Ca2+-activated K +(BKC3) channels and sarcoplasmic reticulum inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) are expressed in a wide variety of cell types, including arterial smooth muscle cells. Here, we studied BKCa channel regulation by IP3 and IP 3Rs in rat and mouse cerebral artery smooth muscle cells. IP 3 activated BKCa channels both in intact cells and in excised inside-out membrane patches. IP3 caused concentration- dependent BKCa channel activation with an apparent dissociation constant (Kd) of ∼4 μM at physiological voltage (-40 mV) and intracellular Ca2+ concentration ( [Ca2+]i; 10 μM). IP3 also caused a leftward-shift in BKCa channel apparent Ca2+ sensitivity and reduced the Kd for free [Ca2+i from ∼20 to 12 μM, but did not alter the slope or maximal Po. BAPTA, a fast Ca2+ buffer, or an elevation in extracellular Ca2+ concentration did not alter IPs-induced BKCa channel activation. Heparin, an IP3R inhibitor, and a monoclonal type 1 IP3R (IP3R1) antibody blocked IP3-induced BKCa channel activation. Adenophostin A, an IP3R agonist, also activated BKCa channels. IP 3 activated BKCa channels in inside-out patches from wild-type (IP3R1+/+) mouse arterial smooth muscle cells, but had no effect on BKCa channels of IP2R1-deficient (IP3R1-/-) mice. Immunofluorescence resonance energy transfer microscopy indicated that IP3R1 is located in close spatial proximity to BKCa α subunits. The IP3R1 monoclonal antibody coimmunoprecipitated IP3R1 and BKCa, channel α and β1 subunits from cerebral arteries. In summary, data indicate that IP3R1 activation elevates BKCa channel apparent Ca2+ sensitivity through local molecular coupling in arterial smooth muscle cells.",
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AU - Leo, Marie Dennis

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AU - Ouyang, Kunfu

AU - Chen, Ju

AU - Jaggar, Jonathan

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