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Hypotension due to Kir6.1 gain-of-function in vascular smooth muscle.

ABSTRACT: KATP channels, assembled from pore-forming (Kir6.1 or Kir6.2) and regulatory (SUR1 or SUR2) subunits, link metabolism to excitability. Loss of Kir6.2 results in hypoglycemia and hyperinsulinemia, whereas loss of Kir6.1 causes Prinzmetal angina-like symptoms in mice. Conversely, overactivity of Kir6.2 induces neonatal diabetes in mice and humans, but consequences of Kir6.1 overactivity are unknown.We generated transgenic mice expressing wild-type (WT), ATP-insensitive Kir6.1 [Gly343Asp] (GD), and ATP-insensitive Kir6.1 [Gly343Asp,Gln53Arg] (GD-QR) subunits, under Cre-recombinase control. Expression was induced in smooth muscle cells by crossing with smooth muscle myosin heavy chain promoter-driven tamoxifen-inducible Cre-recombinase (SMMHC-Cre-ER) mice. Three weeks after tamoxifen induction, we assessed blood pressure in anesthetized and conscious animals, as well as contractility of mesenteric artery smooth muscle and KATP currents in isolated mesenteric artery myocytes. Both systolic and diastolic blood pressures were significantly reduced in GD and GD-QR mice but normal in mice expressing the WT transgene and elevated in Kir6.1 knockout mice as well as in mice expressing dominant-negative Kir6.1 [AAA] in smooth muscle. Contractile response of isolated GD-QR mesenteric arteries was blunted relative to WT controls, but nitroprusside relaxation was unaffected. Basal KATP conductance and pinacidil-activated conductance were elevated in GD but not in WT myocytes.KATP overactivity in vascular muscle can lead directly to reduced vascular contractility and lower blood pressure. We predict that gain of vascular KATP function in humans would lead to a chronic vasodilatory phenotype, as indeed has recently been demonstrated in Cantu syndrome.

SUBMITTER: Li A

PROVIDER: S-EPMC3828800 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

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Hypotension due to Kir6.1 gain-of-function in vascular smooth muscle.

Li Anlong A Knutsen Russell H RH Zhang Haixia H Osei-Owusu Patrick P Moreno-Dominguez Alex A Harter Theresa M TM Uchida Keita K Remedi Maria S MS Dietrich Hans H HH Bernal-Mizrachi Carlos C Blumer Kendall J KJ Mecham Robert P RP Koster Joseph C JC Nichols Colin G CG

Journal of the American Heart Association 20130823 4

<h4>Background</h4>KATP channels, assembled from pore-forming (Kir6.1 or Kir6.2) and regulatory (SUR1 or SUR2) subunits, link metabolism to excitability. Loss of Kir6.2 results in hypoglycemia and hyperinsulinemia, whereas loss of Kir6.1 causes Prinzmetal angina-like symptoms in mice. Conversely, overactivity of Kir6.2 induces neonatal diabetes in mice and humans, but consequences of Kir6.1 overactivity are unknown.<h4>Methods and results</h4>We generated transgenic mice expressing wild-type (WT ...[more]

PMID: 23974906

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Project description:Key pointsSpontaneous contractions are essential for normal lymph transport and these contractions are exquisitely sensitive to the KATP channel activator pinacidil. KATP channel Kir6.1 and SUR2B subunits are expressed in mouse lymphatic smooth muscle (LSM) and form functional KATP channels as verified by electrophysiological techniques. Global deletion of Kir6.1 or SUR2 subunits results in severely impaired lymphatic contractile responses to pinacidil. Smooth muscle-specific expression of Kir6.1 gain-of-function mutant (GoF) subunits results in profound lymphatic contractile dysfunction and LSM hyperpolarization that is partially rescued by the KATP inhibitor glibenclamide. In contrast, lymphatic endothelial-specific expression of Kir6.1 GoF has essentially no effect on lymphatic contractile function. The high sensitivity of LSM to KATP channel GoF offers an explanation for the lymphoedema observed in patients with Cantú syndrome, a disorder caused by gain-of-function mutations in genes encoding Kir6.1 or SUR2, and suggests that glibenclamide may be an appropriate therapeutic agent.AbstractThis study aimed to understand the functional expression of KATP channel subunits in distinct lymphatic cell types, and assess the consequences of altered KATP channel activity on lymphatic pump function. KATP channel subunits Kir6.1 and SUR2B were expressed in mouse lymphatic muscle by PCR, but only Kir6.1 was expressed in lymphatic endothelium. Spontaneous contractions of popliteal lymphatics from wild-type (WT) (C57BL/6J) mice, assessed by pressure myography, were very sensitive to inhibition by the SUR2-specific KATP channel activator pinacidil, which hyperpolarized both mouse and human lymphatic smooth muscle (LSM). In vessels from mice with deletion of Kir6.1 (Kir6.1-/- ) or SUR2 (SUR2[STOP]) subunits, contractile parameters were not significantly different from those of WT vessels, suggesting that basal KATP channel activity in LSM is not an essential component of the lymphatic pacemaker, and does not exert a strong influence over contractile strength. However, these vessels were >100-fold less sensitive than WT vessels to pinacidil. Smooth muscle-specific expression of a Kir6.1 gain-of-function (GoF) subunit resulted in severely impaired lymphatic contractions and hyperpolarized LSM. Membrane potential and contractile activity was partially restored by the KATP channel inhibitor glibenclamide. In contrast, lymphatic endothelium-specific expression of Kir6.1 GoF subunits had negligible effects on lymphatic contraction frequency or amplitude. Our results demonstrate a high sensitivity of lymphatic contractility to KATP channel activators through activation of Kir6.1/SUR2-dependent channels in LSM. In addition, they offer an explanation for the lymphoedema observed in patients with Cantú syndrome, a disorder caused by gain-of-function mutations in genes encoding Kir6.1/SUR2.

Project description:BackgroundRecent studies have demonstrated that the activation of NADPH oxidase 1 (Nox1) plays an important role in the control of reactive oxygen species and their involvement in vascular physiology and pathophysiology. In order to function properly, Nox1 needs to be available in an optimal state, where it is ready to respond appropriately and efficiently to upstream signals. It must also be able to return quickly to this state as soon as the input signal disappears. While Nox1 activation has been discussed extensively in recent years, mechanisms for enzyme disassembly and proper subunit recovery have not received the same attention and therefore require investigation.ResultsWe study the Nox1 system in vascular smooth smucle cells and propose four potential disassembly mechanisms. The analysis consists primarily of large-scale Monte-Carlo simulations whose results are essentially independent of specific parameter values. The computational analysis shows that a specific profile of subunit concentrations is crucial for optimal functioning and responsiveness of the system to input signals. Specifically, free p47(phox) and inactive Rac1 should be dominant under unstimulated resting conditions, and the proteolytic disassembly pathway should have a low flux, as it is relatively inefficient. The computational results also reveal that the optimal design of the three subunit recovery pathways depends on the intracellular settings of the pathway and that the response speeds of key reversible reactions within the pathway are of great importance.ConclusionsOur results provide a systematic basis for understanding the dynamics of Nox1 and yield novel insights into its crucially important disassembly mechanisms. The rigorous comparisons of the relative importance of four potential disassembly pathways demonstrate that disassembly via proteolysis is the least effective mechanism. The relative significance of the other three recovery pathways varies among different scenarios. It is greatly affected by the required response speed of the system and depends critically on appropriate flux balances between forward and reverse reactions. Our findings are predictive and pose novel hypotheses that should be validated with future experiments.

Dataset Information

Hypotension due to Kir6.1 gain-of-function in vascular smooth muscle.

Publications

Hypotension due to Kir6.1 gain-of-function in vascular smooth muscle.

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