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Mechanisms underlying selective coupling of endothelial Ca²⁺ signals with eNOS vs. IK/SK channels in systemic and pulmonary arteries.

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Endothelial cell TRPV4 (TRPV4_EC ) channels exert a dilatory effect on the resting diameter of resistance mesenteric and pulmonary arteries. Functional intermediate- and small-conductance K⁺ (IK and SK) channels and endothelial nitric oxide synthase (eNOS) are present in the endothelium of mesenteric and pulmonary arteries. TRPV4_EC sparklets preferentially couple with IK/SK channels in mesenteric arteries and with eNOS in pulmonary arteries. TRPV4_EC channels co-localize with IK/SK channels in mesenteric arteries but not in pulmonary arteries, which may explain TRPV4_EC -IK/SK channel coupling in mesenteric arteries and its absence in pulmonary arteries. The presence of the nitric oxide-scavenging protein, haemoglobin α, limits TRPV4_EC -eNOS signalling in mesenteric arteries. Spatial proximity of TRPV4_EC channels with eNOS and the absence of haemoglobin α favour TRPV4_EC -eNOS signalling in pulmonary arteries.

Abstract

Spatially localized Ca²⁺ signals activate Ca²⁺ -sensitive intermediate- and small-conductance K⁺ (IK and SK) channels in some vascular beds and endothelial nitric oxide synthase (eNOS) in others. The present study aimed to uncover the signalling organization that determines selective Ca²⁺ signal to vasodilatory target coupling in the endothelium. Resistance-sized mesenteric arteries (MAs) and pulmonary arteries (PAs) were used as prototypes for arteries with predominantly IK/SK channel- and eNOS-dependent vasodilatation, respectively. Ca²⁺ influx signals through endothelial transient receptor potential vanilloid 4 (TRPV4_EC ) channels played an important role in controlling the baseline diameter of both MAs and PAs. TRPV4_EC channel activity was similar in MAs and PAs. However, the TRPV4 channel agonist GSK1016790A (10 nm) selectively activated IK/SK channels in MAs and eNOS in PAs, revealing preferential TRPV4_EC -IK/SK channel coupling in MAs and TRPV4_EC -eNOS coupling in PAs. IK/SK channels co-localized with TRPV4_EC channels at myoendothelial projections (MEPs) in MAs, although they lacked the spatial proximity necessary for their activation by TRPV4_EC channels in PAs. Additionally, the presence of the NO scavenging protein haemoglobin α (Hbα) within nanometer proximity to eNOS limits TRPV4_EC -eNOS signalling in MAs. By contrast, co-localization of TRPV4_EC channels and eNOS at MEPs, and the absence of Hbα, favour TRPV4_EC -eNOS coupling in PAs. Thus, our results reveal that differential spatial organization of signalling elements determines TRPV4_EC -IK/SK vs. TRPV4_EC -eNOS coupling in resistance arteries.

SUBMITTER: Ottolini M

PROVIDER: S-EPMC7484264 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:1. Nine bis-quinolinyl and bis-quinolinium compounds related to dequalinium, and previously shown to block apamin-sensitive small conductance Ca(2+)-activated K(+) channels (SK(Ca)), have been tested for their inhibitory effects on actions mediated by intermediate conductance Ca(2+)-activated K(+) channels (IK(Ca)) in rabbit blood cells. 2. In most experiments, a K(+)-sensitive electrode was employed to monitor the IK(Ca)-mediated net loss of cell K(+) that followed the addition of the Ca(2+) ionophore A23187 (2 microM) to red cells suspended at an haematocrit of 1% in a low K(+) (0.12 - 0.17 mM) solution. The remainder used an optical method based on measuring the reduction in light transmission that occurred on applying A23187 (0.4 or 2 microM) to a very dilute suspension of red cells (haematocrit 0.02%). 3. Of the compounds tested, the most potent IK(Ca) blocker was 1,12 bis[(2-methylquinolin-4-yl)amino]dodecane (UCL 1407) which had an IC(50) of 0.85+/-0.06 microM (mean+/-s.d. mean). 4. The inhibitory action of UCL 1407 and its three most active congeners was characterized by (i) a Hill slope greater than unity, (ii) sensitivity to an increase in external [K(+)], and (iii) a time course of onset that suggested use-dependence. Also, the potency of the nonquaternary compounds tested increased with their predicted lipophilicity. These findings suggested that the IK(Ca) blocking action resembles that of cetiedil rather than of clotrimazole. 5. Some quaternized members of the series were also active. The most potent was the monoquaternary UCL 1440 ((1-[N-[1-(3, 5-dimethoxybenzyl)-2-methylquinolinium-4-yl]amino]-10-[N'-(2-me thylqu inolinium-4yl)amino] decane (trifluoroacetate) which had an IC(50) of 1.8+/-0.1 microM. The corresponding bisquaternary UCL 1438 (1, 10-bis[N-[1-(3,5-dimethoxybenzyl)-2-methylquinolinium-4-yl]amino] decane bis(trifluoroacetate) was almost as active (IC(50) 2.7+/-0.3 microM). 6. A bis-aminoquinolium cyclophane (UCL 1684) had little IK(Ca) blocking action despite its great potency at SK(Ca) channels (IC(50) 4.1+/-0.2 nM). 7. The main outcome is the identification of new intermediate-conductance Ca(2+)-activated K(+) channel blockers with a wide range of IK(Ca)/SK(Ca) selectivities.

Project description:Background and purposeIn small arteries, small conductance Ca²?-activated K? channels (SK(Ca)) and intermediate conductance Ca²?-activated K? channels (IK(Ca)) restricted to the vascular endothelium generate hyperpolarization that underpins the NO- and PGI?-independent, endothelium-derived hyperpolarizing factor response that is the predominate endothelial mechanism for vasodilatation. As neuronal IK(Ca) channels can be negatively regulated by PKA, we investigated whether ?-adrenoceptor stimulation, which signals through cAMP/PKA, might influence endothelial cell hyperpolarization and as a result modify the associated vasodilatation.Experimental approachRat isolated small mesenteric arteries were pressurized to measure vasodilatation and endothelial cell [Ca²?]i , mounted in a wire myograph to measure smooth muscle membrane potential or dispersed into endothelial cell sheets for membrane potential recording.Key resultsIntraluminal perfusion of ?-adrenoceptor agonists inhibited endothelium-dependent dilatation to ACh (1 nM-10 ?M) without modifying the associated changes in endothelial cell [Ca²?]i . The inhibitory effect of ?-adrenoceptor agonists was mimicked by direct activation of adenylyl cyclase with forskolin, blocked by the ?-adrenoceptor antagonists propranolol (non-selective), atenolol (??) or the PKA inhibitor KT-5720, but remained unaffected by ICI 118 551 (??) or glibenclamide (ATP-sensitive K? channels channel blocker). Endothelium-dependent hyperpolarization to ACh was also inhibited by ?-adrenoceptor stimulation in both intact arteries and in endothelial cells sheets. Blocking IK(Ca) {with 1 ?M 1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34)}, but not SK(Ca) (50 nM apamin) channels prevented ?-adrenoceptor agonists from suppressing either hyperpolarization or vasodilatation to ACh.Conclusions and implicationsIn resistance arteries, endothelial cell ??-adrenoceptors link to inhibit endothelium-dependent hyperpolarization and the resulting vasodilatation to ACh. This effect appears to reflect inhibition of endothelial IK(Ca) channels and may be one consequence of raised circulating catecholamines.

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Mechanisms underlying selective coupling of endothelial Ca²⁺ signals with eNOS vs. IK/SK channels in systemic and pulmonary arteries.

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Mechanisms underlying selective coupling of endothelial Ca2+ signals with eNOS vs. IK/SK channels in systemic and pulmonary arteries.

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OmicsDI is part of the ELIXIR infrastructure

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Mechanisms underlying selective coupling of endothelial Ca²⁺ signals with eNOS vs. IK/SK channels in systemic and pulmonary arteries.