Chronic hypoxia limits H<sub>2</sub>O<sub>2</sub>-induced inhibition of ASIC1-dependent store-operated calcium entry in pulmonary arterial smooth muscle

Plomaritas, Danielle; Herbert, Lindsay M.; Yellowhair, Tracylyn R.; Resta, Thomas C.; Bosc, Laura V. González; Walker, Benjimen R.; Jernigan, Nikki L.

doi:10.1152/ajplung.00095.2014

Cited by 28 publications

(35 citation statements)

References 60 publications

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“…We previously determined the specificity of DHE to detect O 2 ·− over H 2 O 2 in this preparation by demonstrating that PEG-superoxide dismutase, but not PEG-catalase decreases DHE fluorescence [15]. Additionally, here we show that DHE does not change in PASMC pre-incubated with increasing doses of H 2 O 2 (Fig 7C).…”

Section: Resultssupporting

confidence: 54%

“…As a consequence of reduced SOD expression and activity, a decrease in H 2 O 2 levels has also been reported following chronic hypoxia exposure [11, 12, 14, 15, 58], as well as in other experimental models of spontaneous PH and in humans with idiopathic PAH [10, 11, 14]. This decrease in H 2 O 2 is thought to contribute to proproliferative and antiapoptotic effects that are, in part, mediated by hypoxia-inducible factor 1α (HIF-1α) and involve a decrease in K v 1.5 channel expression and elevated [Ca 2+ ] i levels [11].…”

Section: Discussionmentioning

confidence: 99%

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Contribution of reactive oxygen species to the pathogenesis of pulmonary arterial hypertension

et al. 2017

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Pulmonary arterial hypertension is associated with a decreased antioxidant capacity. However, neither the contribution of reactive oxygen species to pulmonary vasoconstrictor sensitivity, nor the therapeutic efficacy of antioxidant strategies in this setting are known. We hypothesized that reactive oxygen species play a central role in mediating both vasoconstrictor and arterial remodeling components of severe pulmonary arterial hypertension. We examined the effect of the chemical antioxidant, TEMPOL, on right ventricular systolic pressure, vascular remodeling, and enhanced vasoconstrictor reactivity in both chronic hypoxia and hypoxia/SU5416 rat models of pulmonary hypertension. SU5416 is a vascular endothelial growth factor receptor antagonist and the combination of chronic hypoxia/SU5416 produces a model of severe pulmonary arterial hypertension with vascular plexiform lesions/fibrosis that is not present with chronic hypoxia alone. The major findings from this study are: 1) compared to hypoxia alone, hypoxia/SU5416 exposure caused more severe pulmonary hypertension, right ventricular hypertrophy, adventitial lesion formation, and greater vasoconstrictor sensitivity through a superoxide and Rho kinase-dependent Ca2+ sensitization mechanism. 2) Chronic hypoxia increased medial muscularization and superoxide levels, however there was no effect of SU5416 to augment these responses. 3) Treatment with TEMPOL decreased right ventricular systolic pressure in both hypoxia and hypoxia/SU5416 groups. 4) This effect of TEMPOL was associated with normalization of vasoconstrictor responses, but not arterial remodeling. Rather, medial hypertrophy and adventitial fibrotic lesion formation were more pronounced following chronic TEMPOL treatment in hypoxia/SU5416 rats. Our findings support a major role for reactive oxygen species in mediating enhanced vasoconstrictor reactivity and pulmonary hypertension in both chronic hypoxia and hypoxia/SU5416 rat models, despite a paradoxical effect of antioxidant therapy to exacerbate arterial remodeling in animals with severe pulmonary arterial hypertension in the hypoxia/SU5416 model.

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Section: Resultssupporting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Contribution of reactive oxygen species to the pathogenesis of pulmonary arterial hypertension

et al. 2017

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“…We have shown the same alteration in the O 2 ·Ϫ -to-H 2 O 2 ratio in pulmonary arteries of SOD1 KO mice (62) and rats exposed to CH (59). Furthermore, it has been shown that ET-1 reduces SOD activity and expression in the systemic vasculature (15,16).…”

Section: Discussionsupporting

confidence: 56%

Mechanisms of NFATc3 activation by increased superoxide and reduced hydrogen peroxide in pulmonary arterial smooth muscle

Ramiro-Diaz

Giermakowska

Weaver

et al. 2014

American Journal of Physiology-Cell Physiology

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·Ϫ ) and decreased H2O2 levels in pulmonary arteries of chronic hypoxia-exposed wild-type and normoxic superoxide dismutase 1 (SOD1) knockout mice. We also showed that this reciprocal change in O 2 ·Ϫ and H2O2 is associated with elevated activity of nuclear factor of activated T cells isoform c3 (NFATc3) in pulmonary arterial smooth muscle cells (PASMC). This suggests that an imbalance in reactive oxygen species levels is required for NFATc3 activation. However, how such imbalance activates NFATc3 is unknown. This study evaluated the importance of O 2 ·Ϫ and H2O2 in the regulation of NFATc3 activity. We tested the hypothesis that an increase in O2 ·Ϫ enhances actin cytoskeleton dynamics and a decrease in H2O2 enhances intracellular Ca 2ϩ concentration, contributing to NFATc3 nuclear import and activation in PASMC. We demonstrate that, in PASMC, endothelin-1 increases O 2 ·Ϫ while decreasing H2O2 production through the decrease in SOD1 activity without affecting SOD protein levels. We further demonstrate that O 2 ·Ϫ promotes, while H2O2 inhibits, NFATc3 activation in PASMC. Additionally, increased O 2 ·Ϫ -to-H2O2 ratio activates NFATc3, even in the absence of a G q protein-coupled receptor agonist. Furthermore, O2·Ϫ -dependent actin polymerization and low intracellular H2O2 concentration-dependent increases in intracellular Ca 2ϩ concentration contribute to NFATc3 activation. Together, these studies define important and novel regulatory mechanisms of NFATc3 activation in PASMC by reactive oxygen species.superoxide; hydrogen peroxide; NFATc3; endothelin-1; actin cytoskeleton; calcium NUCLEAR FACTOR OF ACTIVATED T cells (NFAT) isoform c3 (NFATc3) belongs to a family of four transcription factors (NFATc1, NFATc2, NFATc3, and NFATc4) that share the property of Ca 2ϩ /calcineurin-dependent nuclear translocation (reviewed in Ref. 35). We previously demonstrated that NFATc3 is required for chronic hypoxia (CH)-induced pulmonary arterial remodeling and pulmonary hypertension in mice (10, 25). Pulmonary hypertension is associated with polycythemia, arterial remodeling, increased vascular contractility, augmented concentrations of circulating endothelin-1 (ET-1), and elevated reactive oxygen species (ROS) (1,9,19,24,28,32,36,55 (2): SOD1 and extracellular SOD3 (Cu,Zn-SOD) and mitochondrial SOD2 (Mn-SOD); SOD1, the predominant cytosolic isoform (4), is also present in the mitochondrial intermembrane space (42). We recently showed that SOD1 knockout (KO) mice develop NFAT-dependent spontaneous pulmonary hypertension (62). CH-exposed wild-type and normoxic SOD1 KO mice have increased O 2 ·Ϫ and decreased H 2 O 2 levels in pulmonary arteries. This O 2 ·Ϫ -H 2 O 2 imbalance is associated with elevated NFATc3 activity in pulmonary arterial smooth muscle cells (PASMC) (62). Similarly, in rats exposed to CH, we recently showed decreased pulmonary arterial SOD1 activity/expression, leading to increased O 2 ·Ϫ and decreased H 2 O 2 production (59), as previously reported in newborn piglets (26). These findings suggest that an imbala...

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“…PASMC were superfused (5 ]i) were determined upon repletion of HEPES-based PSS containing 1.8 mM CaCl2 in the continued presence of diltiazem and CPA. Area under the curve was calculated as previously described (36). Experiments were conducted in the absence or presence of the PICK1 inhibitor FSC231 (50 M; Calbiochem), the ASIC1 inhibitor PcTX1 (20 nM; Phoenix Peptides), or in PASMC transfected with the negative or PICK1 shRNA.…”

Section: Role Of Pick1 To Regulate Asic1-dependent Socementioning

confidence: 99%

PICK1/calcineurin suppress ASIC1-mediated Ca²⁺ entry in rat pulmonary arterial smooth muscle cells

Herbert

Nitta

Yellowhair

et al. 2016

American Journal of Physiology-Cell Physiology

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Herbert LM, Nitta CH, Yellowhair TR, Browning C, Gonzalez Bosc LV, Resta TC, Jernigan NL. PICK1/calcineurin suppress ASIC1-mediated Ca 2ϩ entry in rat pulmonary arterial smooth muscle cells. Am J Physiol Cell Physiol 310: C390 -C400, 2016. First published December 23, 2015; doi:10.1152/ajpcell.00091.2015.-Acidsensing ion channel 1 (ASIC1) contributes to Ca 2ϩ influx and contraction in pulmonary arterial smooth muscle cells (PASMC). ASIC1 binds the PDZ (PSD-95/Dlg/ZO-1) domain of the protein interacting with C kinase 1 (PICK1), and this interaction is important for the subcellular localization and/or activity of ASIC1. Therefore, we first hypothesized that PICK1 facilitates ASIC1-dependent Ca 2ϩ influx in PASMC by promoting plasma membrane localization. Using Duolink to determine protein-protein interactions and a biotinylation assay to assess membrane localization, we demonstrated that the PICK1 PDZ domain inhibitor FSC231 diminished the colocalization of PICK1 and ASIC1 but did not limit ASIC1 plasma membrane localization. Although stimulation of store-operated Ca 2ϩ entry (SOCE) greatly enhanced colocalization between ASIC1 and PICK1, both FSC231 and shRNA knockdown of PICK1 largely augmented SOCE. These data suggest PICK1 imparts a basal inhibitory effect on ASIC1 Ca 2ϩ entry in PASMC and led to an alternative hypothesis that PICK1 facilitates the interaction between ASIC1 and negative intracellular modulators, namely PKC and/or the calcium-calmodulin-activated phosphatase calcineurin. FSC231 limited PKC-mediated inhibition of SOCE, supporting a potential role for PICK1 in this response. Additionally, we found PICK1 inhibits ASIC1-mediated SOCE through an effect of calcineurin to dephosphorylate the channel. Furthermore, it appears PICK1/calcineurin-mediated regulation of SOCE opposes PKA phosphorylation and activation of ASIC1. Together our data suggest PKA and PICK1/calcineurin differentially regulate ASIC1-mediated SOCE and these modulatory complexes are important in determining downstream Ca 2ϩ signaling.store-operated calcium entry; FSC231; DEG/ENaC; Duolink; PKA; PKC ACID-SENSING ION CHANNELS (ASIC) belong to the degenerin/ epithelial sodium channel (DEG/ENaC) superfamily, which includes several amiloride-sensitive cation channels. Significant progress has been made in understanding the structure and function of ASIC in the nervous system; however, several questions remain regarding their physiological importance in other tissues. There is an emerging role for both ENaC and ASIC in vascular smooth muscle and endothelial cells from a variety of vascular beds (8, 10 -12, 19, 20, 25, 33, 46). Consistent with a physiological role of ASIC in the vasculature, our laboratory has recently shown that ASIC1 is an important facilitator of G protein-coupled receptor signaling via store-operated Ca 2ϩ entry (SOCE) in pulmonary arterial smooth muscle cells (PASMC) (21, 22). Furthermore, ASIC1-mediated Ca 2ϩ entry in PASMC appears to be an important constituent of both the active vasoconstrictor and vascular remodelin...

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Chronic hypoxia limits H₂O₂-induced inhibition of ASIC1-dependent store-operated calcium entry in pulmonary arterial smooth muscle

Cited by 28 publications

References 60 publications

Contribution of reactive oxygen species to the pathogenesis of pulmonary arterial hypertension

Contribution of reactive oxygen species to the pathogenesis of pulmonary arterial hypertension

Mechanisms of NFATc3 activation by increased superoxide and reduced hydrogen peroxide in pulmonary arterial smooth muscle

PICK1/calcineurin suppress ASIC1-mediated Ca²⁺ entry in rat pulmonary arterial smooth muscle cells

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Chronic hypoxia limits H2O2-induced inhibition of ASIC1-dependent store-operated calcium entry in pulmonary arterial smooth muscle

Cited by 28 publications

References 60 publications

Contribution of reactive oxygen species to the pathogenesis of pulmonary arterial hypertension

Contribution of reactive oxygen species to the pathogenesis of pulmonary arterial hypertension

Mechanisms of NFATc3 activation by increased superoxide and reduced hydrogen peroxide in pulmonary arterial smooth muscle

PICK1/calcineurin suppress ASIC1-mediated Ca2+ entry in rat pulmonary arterial smooth muscle cells

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Chronic hypoxia limits H₂O₂-induced inhibition of ASIC1-dependent store-operated calcium entry in pulmonary arterial smooth muscle

PICK1/calcineurin suppress ASIC1-mediated Ca²⁺ entry in rat pulmonary arterial smooth muscle cells