Increases in mitochondrial reactive oxygen species trigger hypoxia-induced calcium responses in pulmonary artery smooth muscle cells
Abstract
Mitochondria have been implicated as a potential site of O(2) sensing underlying hypoxic pulmonary vasoconstriction (HPV), but 2 disparate models have been proposed to explain their reaction to hypoxia. One model proposes that hypoxia-induced increases in mitochondrial reactive oxygen species (ROS) generation activate HPV through an oxidant-signaling pathway, whereas the other proposes that HPV is a result of decreased oxidant signaling. In an attempt to resolve this debate, we use a novel, ratiometric, redox-sensitive fluorescence resonance energy transfer (HSP-FRET) probe, in concert with measurements of reduced/oxidized glutathione (GSH/GSSG), to assess cytosolic redox responses in cultured pulmonary artery smooth muscle cells (PASMCs). Superfusion of PASMCs with hypoxic media increases the HSP-FRET ratio and decreases GSH/GSSG, indicating an increase in oxidant stress. The antioxidants pyrrolidinedithiocarbamate and N-acetyl-l-cysteine attenuated this response, as well as the hypoxia-induced increases in cytosolic calcium ([Ca(2+)](i)), assessed by the Ca(2+)-sensitive FRET sensor YC2.3. Adenoviral overexpression of glutathione peroxidase or cytosolic or mitochondrial catalase attenuated the hypoxia-induced increase in ROS signaling and [Ca(2+)](i). Adenoviral overexpression of cytosolic Cu, Zn-superoxide dismutase (SOD-I) had no effect on the hypoxia-induced increase in ROS signaling and [Ca(2+)](i), whereas mitochondrial matrix-targeted Mn-SOD (SOD-II) augmented [Ca(2+)](i). The mitochondrial inhibitor myxothiazol attenuated the hypoxia-induced changes in the ROS signaling and [Ca(2+)](i), whereas cyanide augmented the increase in [Ca(2+)](i). Finally, simultaneous measurement of ROS and Ca(2+) signaling in the same cell revealed that the initial increase in these 2 signals could not be distinguished temporally. These results demonstrate that hypoxia triggers increases in PASMC [Ca(2+)](i) by augmenting ROS signaling from the mitochondria.
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Hypoxia leads to an increase in mitochondrial production of ROS.
“Paradoxically, hypoxia leads to an increase in mitochondrial production of ROS.”
From Mitochondrial reactive oxygen species regulate cellular signaling and dictate biological outcomes (2010) · cites this paper as [14]
H2O2 emission from mitochondria during hypoxia is a central upstream regulator of many cellular responses to hypoxia.
“Current evidence suggests that this H2O2 emission from mitochondria during hypoxia is a central upstream regulator of many of the cellular responses to hypoxia 8–14.”
From Mitochondrial reactive oxygen species regulate cellular signaling and dictate biological outcomes (2010) · cites this paper as [14]
The increase in calcium and subsequent pulmonary artery vasoconstriction are both dependent on hypoxic mitochondrial ROS production.
“This increase in calcium and subsequent pulmonary artery vasoconstriction are both dependent on hypoxic mitochondrial ROS production 9, 14.”
From Mitochondrial reactive oxygen species regulate cellular signaling and dictate biological outcomes (2010) · cites this paper as [14]
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- Authors
- Waypa, G. B., Guzy, R., Mungai, P. T., Mack, M. M., Marks, J. D., Roe, M. W., & Schumacker, P. T.
- Affiliations
- Northwestern University (US)
- Venue
- Circulation Research
- Published
- 2006-10-27
- Volume
- 99
- Issue
- 9
- Pages
- 970-978
- ISSN
- 0009-7330 (ISSN-L)
- 1524-4571
- OpenAlex
- W2111983287
- Cited by
- 191
- References
- 46
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 4 · 2023: 7 · 2022: 7 · 2021: 13 · 2020: 3 · 2019: 6 · 2018: 15 · 2017: 16 · 2016: 6 · 2015: 5 · 2014: 14 · 2013: 10 · 2012: 11
- FWCI
- 11.49
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Circulation Research (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (R01HL066315, HL079650, HL66315, R01HL035440, R01HL079650, HL35440)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK-63493, R01DK063493)
- National Institute of Neurological Disorders and Stroke (R01NS038547)
- Topics
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Pulmonary Hypertension Research and Treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Keywords
- hypoxic pulmonary vasoconstriction, mitochondrial reactive oxygen species, pulmonary artery smooth muscle cells, cytosolic calcium signaling, oxidant signaling, glutathione redox state, superoxide dismutase, catalase, glutathione peroxidase, myxothiazol, oxygen sensing
- MeSH terms
- Animals, Calcium, Cytosol, Glutathione Peroxidase, Mitochondria, Muscle, Smooth, Vascular, Oxidation-Reduction, Pulmonary Artery, Vasoconstriction, Signal Transduction, Cell Hypoxia, Reactive Oxygen Species, Calcium Signaling, Fluorescence Resonance Energy Transfer, Myocytes, Smooth Muscle, Rats
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