Dataset Information

A Role of BK Channel in Regulation of Ca2+ Channel in Ventricular Myocytes by Substrate Stiffness.

ABSTRACT: Substrate stiffness is crucial for diverse cell functions, but the mechanisms conferring cells with mechanosensitivity are still elusive. By tailoring substrate stiffness with 10-fold difference, we showed that L-type voltage-gated Ca2+ channel current density was greater in chick ventricular myocytes cultured on the stiff substrate than on the soft substrate. Blockage of the BK channel increased the Ca2+ current density on the soft substrate and consequently eliminated substrate stiffness regulation of the Ca2+ channel. The expression of the BK channel, including the STREX-containing ?-subunit that forms stretch-activated BK channel in myocytes and the BK channel function in myocytes (and also in HEK293 cells heterologously expressing STREX-containing ?- and ?1-subunits) was reduced in cells cultured on the stiff substrate. Furthermore, in HEK293 cells coexpressing the cardiac CaV1.2 channel and STREX-containing BK channel, the Ca2+ current density was greater in cells on the stiff substrate, which was not observed in cells expressing the CaV1.2 channel alone or coexpressing with the STREX-deleted BK channel. These results provide strong evidence to show that the stretch-activated BK channel plays a key role in functional regulation of cardiac voltage-gated Ca2+ channel by substrate stiffness, revealing, to our knowledge, a novel mechanosensing mechanism in ventricular myocytes.

SUBMITTER: Zhao H

PROVIDER: S-EPMC5389963 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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A Role of BK Channel in Regulation of Ca2+ Channel in Ventricular Myocytes by Substrate Stiffness.

Zhao Hucheng H Yu Yang Y Wu Xiaoan X Liu Sisi S Liu Bailin B Du Jing J Li Bo B Jiang Linhua L Feng Xiqiao X

Biophysical journal 20170401 7

Substrate stiffness is crucial for diverse cell functions, but the mechanisms conferring cells with mechanosensitivity are still elusive. By tailoring substrate stiffness with 10-fold difference, we showed that L-type voltage-gated Ca2+ channel current density was greater in chick ventricular myocytes cultured on the stiff substrate than on the soft substrate. Blockage of the BK channel increased the Ca2+ current density on the soft substrate and consequently eliminated sub ...[more]

PMID: 28402883

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Project description:Backgrounds/aimMale and female hearts have many structural and functional differences. Here, we investigated the role of estrogen (E2) in the mechanisms of sex differences in contraction through the cAMP-L-type Ca2+channel pathway in adult mice left ventricular (LV) apical myocytes at basal and stress state.MethodsIsolated LV apical myocytes from male, female (Sham) and ovariectomised mice (OVX) were used to investigate contractility, Ca2+ transients and L-type Ca2+ channel (LTCC) function. The levels of β2AR, intracellular cAMP, phosphodiesterase (PDE 3 and PDE 4), RyR2, PLB, SLN, and SERCA2a were compared among the experimental groups.ResultsWe found that (1) intracellular cAMP, ICaL density, contraction and Ca2+ transient amplitudes were larger in Sham and OVX + E2 myocytes compared to male and OVX. (2) The mRNA expression of PDE 3 and 4 were lower in Sham and OVX + E2 groups compared with male and OVX groups. Treatment of myocytes with IBMX (100 μM) increased contraction and Ca2+ transient amplitude in both sexes and canceled differences between them. (3) β2AR-mediated stress decreased cAMP concentration and peak contraction and Ca2+ transient amplitude only in male and OVX groups but not in Sham or OVX + E2 groups suggesting a cardioprotective role of E2 in female mice. (4) Pretreatment of OVX myocytes with GPR30 antagonist G15 (100 nM) abolished the effects of E2, but ERα and ERβ antagonist ICI 182,780 (1 μM) did not. Moreover, activation of GPR30 with G1 (100 nM) replicated the effects of E2 on cAMP, contraction and Ca2+ transient amplitudes suggesting that the acute effects of E2 were mediated by GPR30 via non-genomic signaling. (5) mRNA expression of RyR2 was higher in myocytes from Sham than those of male while PLB and SLN were higher in male than Sham but no sex differences were observed in the mRNA of SERCA2a.ConclusionCollectively, these results demonstrate that E2 modulates the expression of genes related to the cAMP-LTCC pathway and contributes to sex differences in cardiac contraction and responses to stress. We also show that estrogen confers cardioprotection against cardiac stress by non-genomic acute signaling via GPR30.

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Dataset Information

A Role of BK Channel in Regulation of Ca2+ Channel in Ventricular Myocytes by Substrate Stiffness.

Publications

A Role of BK Channel in Regulation of Ca<sup>2+</sup> Channel in Ventricular Myocytes by Substrate Stiffness.

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