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A junctophilin-caveolin interaction enables efficient coupling between ryanodine receptors and BK_Ca channels in the Ca²⁺ microdomain of vascular smooth muscle.

ABSTRACT: Functional coupling between large-conductance Ca²⁺-activated K⁺ (BK_Ca) channels in the plasma membrane (PM) and ryanodine receptors (RyRs) in the sarcoplasmic reticulum (SR) is an essential mechanism for regulating mechanical force in most smooth muscle (SM) tissues. Spontaneous Ca²⁺ release through RyRs (Ca²⁺ sparks) and subsequent BK_Ca channel activation occur within the PM-SR junctional sites. We report here that a molecular interaction of caveolin-1 (Cav1), a caveola-forming protein, with junctophilin-2 (JP2), a bridging protein between PM and SR, positions BK_Ca channels near RyRs in SM cells (SMCs) and thereby contributes to the formation of a molecular complex essential for Ca²⁺ microdomain function. Approximately half of all Ca²⁺ sparks occurred within a close distance (<400 nm) from fluorescently labeled JP2 or Cav1 particles, when they were moderately expressed in primary SMCs from mouse mesenteric artery. The removal of caveolae by genetic Cav1 ablation or methyl-β-cyclodextrin treatments significantly reduced coupling efficiency between Ca²⁺ sparks and BK_Ca channel activity in SMCs, an effect also observed after JP2 knockdown in SMCs. A 20-amino acid-long region in JP2 appeared to be essential for the observed JP2-Cav1 interaction, and we also observed an interaction between JP2 and the BK_Ca channel. It can be concluded that the JP2-Cav1 interaction provides a structural and functional basis for the Ca²⁺ microdomain at PM-SR junctions and mediates cross-talk between RyRs and BK_Ca channels, converts local Ca²⁺ sparks into membrane hyperpolarization, and contributes to stabilizing resting tone in SMCs.

SUBMITTER: Saeki T

PROVIDER: S-EPMC6721949 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Mammalian circadian (24 h) rhythms are timed by the pattern of spontaneous action potential firing in the suprachiasmatic nucleus (SCN). This oscillation in firing is produced through circadian regulation of several membrane currents, including large-conductance Ca2+- and voltage-activated K+ (BK) and L-type Ca2+ channel (LTCC) currents. During the day steady-state BK currents depend mostly on LTCCs for activation, whereas at night they depend predominantly on ryanodine receptors (RyRs). However, the contribution of these Ca2+ channels to BK channel activation during action potential firing has not been thoroughly investigated. In this study, we used a pharmacological approach to determine that both LTCCs and RyRs contribute to the baseline membrane potential of SCN action potential waveforms, as well as action potential-evoked BK current, during the day and night, respectively. Since the baseline membrane potential is a major determinant of circadian firing rate, we focused on the LTCCs contributing to low voltage activation of BK channels during the subthreshold phase. For these experiments, two LTCC subtypes found in SCN (CaV1.2 and CaV1.3) were coexpressed with BK channels in heterologous cells, where their differential contributions could be separately measured. CaV1.3 channels produced currents that were shifted to more hyperpolarized potentials compared with CaV1.2, resulting in increased subthreshold Ca2+ and BK currents during an action potential command. These results show that although multiple Ca2+ sources in SCN can contribute to the activation of BK current during an action potential, specific BK-CaV1.3 partnerships may optimize the subthreshold BK current activation that is critical for firing rate regulation.NEW & NOTEWORTHY BK K+ channels are important regulators of firing. Although Ca2+ channels are required for their activation in excitable cells, it is not well understood how BK channels activate using these Ca2+ sources during an action potential. This study demonstrates the differences in BK current activated by CaV1.2 and CaV1.3 channels in clock neurons and heterologous cells. The results define how specific ion channel partnerships can be engaged during distinct phases of the action potential.

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A junctophilin-caveolin interaction enables efficient coupling between ryanodine receptors and BK_Ca channels in the Ca²⁺ microdomain of vascular smooth muscle.

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Dataset Information

A junctophilin-caveolin interaction enables efficient coupling between ryanodine receptors and BKCa channels in the Ca2+ microdomain of vascular smooth muscle.

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OmicsDI is part of the ELIXIR infrastructure

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A junctophilin-caveolin interaction enables efficient coupling between ryanodine receptors and BK_Ca channels in the Ca²⁺ microdomain of vascular smooth muscle.