Research ArticlesOpen Access

Quantitative optical measurement of mitochondrial superoxide dynamics in pulmonary artery endothelial cells

Zahra Ghanian

Department of Electrical Engineering, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin, USA

Search for more papers by this author

Girija Ganesh Konduri

http://orcid.org/0000-0002-3602-1538

Department of Pediatrics, Division of Neonatology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

Co-Senior Authors.

Search for more papers by this author

Said Halim Audi

Department of Biomedical Engineering, Marquette University, Milwaukee, Wisconsin, USA

Search for more papers by this author

Amadou K. S. Camara

http://orcid.org/0000-0002-7652-1414

Department of Anesthesiology and Anesthesia Research, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

Co-Senior Authors.

Search for more papers by this author

, and

Mahsa Ranji

Department of Electrical Engineering, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin, USA

E-mail Address: ranji@uwm.edu

Corresponding author.

Co-Senior Authors.

Search for more papers by this author

https://doi.org/10.1142/S1793545817500183Cited by:9 (Source: Crossref)

Abstract

Reactive oxygen species (ROS) play a vital role in cell signaling and redox regulation, but when present in excess, lead to numerous pathologies. Detailed quantitative characterization of mitochondrial superoxide anion (O $_{2}^{• -})$ production in fetal pulmonary artery endothelia cells (PAECs) has never been reported. The aim of this study is to assess mitochondrial O $_{2}^{• -}$ production in cultured PAECs over time using a novel quantitative optical approach. The rate, the sources, and the dynamics of O $_{2}^{• -}$ production were assessed using targeted metabolic modulators of the mitochondrial electron transport chain (ETC) complexes, specifically an uncoupler and inhibitors of the various ETC complexes, and inhibitors of extra-mitochondrial sources of O $_{2}^{• -}$ . After stabilization, the cells were loaded with nanomolar mitochondrial-targeted hydroethidine (Mito-HE, MitoSOX) online during the experiment without washout of the residual dye. Time-lapse fluorescence microscopy was used to monitor the dynamic changes in O $_{2}^{• -}$ fluorescence intensity over time in PAECs. The transient behaviors of the fluorescence time course showed exponential increases in the rate of O $_{2}^{• -}$ production in the presence of the ETC uncoupler or inhibitors. The most dramatic and the fastest increase in O $_{2}^{• -}$ production was observed when the cells were treated with the uncoupling agent, PCP. We also showed that only the complex IV inhibitor, KCN, attenuated the marked surge in O $_{2}^{• -}$ production induced by PCP. The results showed that mitochondrial respiratory complexes I, III and IV are sources of O $_{2}^{• -}$ production in PAECs, and a new observation that ROS production during uncoupling of mitochondrial respiration is mediated in part via complex IV. This novel method can be applied in other studies that examine ROS production under stress condition and during ROS-mediated injuries in vitro.

Keywords:

References

1. A. P. West, G. S. Shadel, S. Ghosh, “Mitochondria in innate immune responses,” Nat. Rev. Immunol. 11, 489–402 (2011). Web of Science, Google Scholar
2. J. St-Pierre, J. A. Buckingham, S. J. Roebuck, M. D. Brand, “Topology of superoxide production from different sites in the mitochondrial electron transport chain,” J. Biolo. Chem. 277, 44784–44790 (2002). Web of Science, Google Scholar
3. B. Chance, H. Sies, A. Boveris, “Hydroperoxide metabolism in mammalian organs,” Physiol. Rev. 59, 527–605 (1979). Web of Science, Google Scholar
4. D. F. Stowe, A. K. S. Camara, “Mitochondrial Reactive Oxygen Species Production in Excitable Cells: Modulators of Mitochondrial and Cell Function,” Antioxid. Redox Signal. 11, 1373–1414 (2009). Web of Science, Google Scholar
5. A. Boveris, B. Chance, “The mitochondrial generation of hydrogen peroxide. General properties and effect of hyperbaric oxygen,” Biochem. J. 134, 707–716 (1973). Web of Science, Google Scholar
6. A. K. S. Camara, E. J. Lesnefsky, D. F. Stowe, “Potential Therapeutic Benefits of Strategies Directed to Mitochondria,” Antioxid. Redox Signal. 13, 279–347 (2010). Web of Science, Google Scholar
7. A. K. S. Camara, M. Bienengraeber, D. F. Stowe, “Mitochondrial approaches to protect against cardiac ischemia and reperfusion injury,” Front. Physiol. 2(13), 1–34 (2011). Google Scholar
8. R. M. Touyz, “Reactive oxygen species and angiotensin II signaling in vascular cells - implications in cardiovascular disease,” Braz. J. Med. Biol. Res. 37, 1263–1273 (2004). Web of Science, Google Scholar
9. F. Q. Schafer, G. R. Buettner, “Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple,” Free Radi. Biol. Med. 30, 1191–1212 (2001). Web of Science, Google Scholar
10. K. Nagata, Y. Iwasaki, T. Yamada, T. Yuba, K. Kono, S. Hosogi et al., “Overexpression of manganese superoxide dismutase by N-acetylcysteine in hyperoxic lung injury,” Respir. Med. 101, 800–807 (2007). Web of Science, Google Scholar
11. E. Gitto, R. J. Reiter, M. Karbownik, X. T. Dun, I. Barberi, “Respiratory distress syndrome in the newborn: Role of oxidative stress,” Intensive Care Med. 27, 1116–1123 (2001). Web of Science, Google Scholar
12. M. E. Wearden, U. T. Brunk, A. Terman, J. W. Eaton, “Mitochondria: Potential importance in hyperoxic lung injury,” Pediatr. Res. 47, 380a–380a (2000). Web of Science, Google Scholar
13. O. D. Saugstad, “Bronchopulmonary dysplasia-oxidative stress and antioxidants,” Semin. Neonatol. 8, 39–49 (2003). Google Scholar
14. N. G. Bazan, V. Colangelo, W. J. Lukiw, “Prostaglandins and other lipid mediators in Alzheimer’s disease,” Prostaglandins Other Lipid Mediat. 68–69, 197–210 (2002). Web of Science, Google Scholar
15. H. L. Hsieh, C. M. Yang, “Role of redox signaling in neuroinflammation and neurodegenerative diseases,” Biomed. Res. Int. 2013, Article ID 484613, 18 pages (2013). Web of Science, Google Scholar
16. J. F. Turrens, “Mitochondrial formation of reactive oxygen species,” J. Physiol.-London 552, 335–344 (2003). Web of Science, Google Scholar
17. V. Sampath, A. C. Radish, A. L. Eis, K. Broniowska, N. Hogg, G. G. Konduri, “Attenuation of lipopolysaccharide-induced oxidative stress and apoptosis in fetal pulmonary artery endothelial cells by hypoxia,” Free Radic. Biol. Med. 46, 663–671 (2009). Web of Science, Google Scholar
18. K. N. Farrow, S. Wedgwood, K. J. Lee, L. Czech, S. F. Gugino, S. Lakshminrusimha et al., “Mitochondrial oxidant stress increases PDE5 activity in persistent pulmonary hypertension of the newborn,” Respir. Physiol. Neurobiol. 174, 272–281 (2010). Web of Science, Google Scholar
19. S. Lakshminrusimha, J. A. Russell, R. H. Steinhorn, D. D. Swartz, R. M. Ryan, S. F. Gugino et al., “Pulmonary Hemodynamics in neonatal lambs resuscitated with 21%, 50%, and 100% oxygen,” Pediatr. Res. 62, 313–318 (2007). Web of Science, Google Scholar
20. P. Mukhopadhyay, M. Rajesh, G. Hasko, B. J. Hawkins, M. Madesh, P. Pacher, “Simultaneous detection of apoptosis and mitochondrial superoxide production in live cells by flow cytometry and confocal microscopy,” Nat. Protoc. 2, 2295–2301 (2007). Web of Science, Google Scholar
21. K. M. Robinson, M. S. Janes, M. Pehar, J. S. Monette, M. F. Ross, T. M. Hagen et al., “Selective fluorescent imaging of superoxide in vivo using ethidium-based probes,” Proc. Nat. Acad. Sci. USA 103, 15038–15043 (2006). Web of Science, Google Scholar
22. H. R. Rezvani, N. Ali, A. Taieb, H. de Verneuil, F. Mazurier, “Hypoxia-inducible factor-1alpha, a key factor in skin physiology and pathophysiology,” J. Invest. Dermatol. 132, S31–S31 (2012). Web of Science, Google Scholar
23. M. Pehar, M. R. Vargas, K. M. Robinson, P. Cassina, P. J. Diaz-Amarilla, T. M. Hagen et al., “Mitochondrial superoxide production and nuclear factor erythroid 2-related factor 2 activation in p75 neurotrophin receptor-induced motor neuron apoptosis,” J. Neurosci. 27, 7777–7785 (2007). Web of Science, Google Scholar
24. P. Mukhopadhyay, M. Rajesh, K. Yoshihiro, G. Hasko, P. Pacher, “Simple quantitative detection of mitochondrial superoxide production in live cells,” Biochem. Biophys. Res. Commun. 358, 203–208 (2007). Web of Science, Google Scholar
25. B. J. Hawkins, M. Madesh, C. J. Kirkpatrick, A. B. Fisher, “Superoxide flux in endothelial cells via the chloride channel-3 mediates intracellular signaling,” Mol. Biol. Cell. 18, 2002–2012 (2007). Web of Science, Google Scholar
26. A. De Pauw, S. Demine, S. Tejerina, M. Dieu, E. Delaive, A. Kel et al., “Mild mitochondrial uncoupling does not affect mitochondrial biogenesis but downregulates pyruvate carboxylase in adipocytes: Role for triglyceride content reduction,” Am. J. Physiol.-Endocrinol. Metabol. 302, E1123–E1141 (2012). Web of Science, Google Scholar
27. A. Y. Abramov, A. Scorziello, M. R. Duchen, “Three distinct mechanisms generate oxygen free radicals in neurons and contribute to cell death during anoxia and reoxygenation,” J. Neurosci. 27, 1129–1138 (2007). Web of Science, Google Scholar
28. M. C. Zimmennan, L. W. Oberley, S. W. Flanagan, “Mutant SOD1-induced neuronal toxicity is mediated by increased mitochondrial superoxide levels,” J. Neurochem. 102, 609–618 (2007). Web of Science, Google Scholar
29. J. Fauconnier, D. C. Andersson, S. J. Zhang, J. T. Lanner, R. Wibom, A. Katz et al., “Effects of palmitate on Ca2 $+$ handling in adult control and ob/ob cardiomyocytes,” Diabetes 56, 1136–1142 (2007). Web of Science, Google Scholar
30. A. Iuso, S. Scacco, C. Piccoli, F. Bellomo, V. Petruzzella, R. Trentadue et al., “Dysfunctions of cellular oxidative metabolism in patients with mutations in the NDUFS1 and NDUFS4 genes of complex I,” J. Biol. Chem. 281, 10374–10380 (2006). Web of Science, Google Scholar
31. M. Rajesh, P. Mukhopadhyay, S. Batkai, G. Hasko, L. Liaudet, V. R. Drel et al., “Cannabidiol attenuates high glucose-induced endothelial cell inflammatory response and barrier disruption,” Am. J. Physiol.-Heart Circul. Physiol. 293, H610–H619 (2007). Web of Science, Google Scholar
32. G. G. Konduri, J. S. Ou, Y. Shi, K. A. Pritchard, “Decreased association of HSP90 impairs endothelial nitric oxide synthase in fetal lambs with persistent pulmonary hypertension,” Am. J. Physiol.-Heart Circul. Physiol. 285, H204–H211 (2003). Web of Science, Google Scholar
33. S. Lakshminrusimha, J. A. Russell, R. H. Steinhorn, R. M. Ryan, S. F. Gugino, F. C. Morin 3rd, et al., “Pulmonary arterial contractility in neonatal lambs increases with 100% oxygen resuscitation,” Pediatr. Res. 59, 137–141 (2006). Web of Science, Google Scholar
34. R. A. J. Smith, R. C. Hartley, M. P. Murphy, “Mitochondria-Targeted Small Molecule Therapeutics and Probes,” Antioxid. Redox Signal 15, 3021–3038 (2011). Web of Science, Google Scholar
35. K. M. Robinson, M. S. Janes, J. S. Beckman, “The selective detection of mitochondrial superoxide by live cell imaging,” Nat. Protoc. 3, 941–947 (2008). Web of Science, Google Scholar
36. Y. B. Liu, D. R. Schubert, “The specificity of neuroprotection by antioxidants” J. Biomed. Sci. 16, 98–109 (2009). Web of Science, Google Scholar
37. S. Bolte, F. P. Cordelieres, “A guided tour into subcellular colocalization analysis in light microscopy,” J. Microsc.-Oxford, 224, 213–232 (2006). Web of Science, Google Scholar
38. W. Strober, “Trypan Blue Exclusion Test of Cell Viability,” Curr. Protoc. Immunol. 111, A3 B 1–3 (2015). Google Scholar
39. Z. Ghanian, K. Staniszewski, N. Jamali, R. Sepehr, S. Wang, C. M. Sorenson et al., “Quantitative assessment of retinopathy using multi-parameter image analysis,” J. Med. Signals Sens. 6, 71–80 (2016). Web of Science, Google Scholar
40. H. R. Rezvani, S. Dedieu, S. North, F. Belloc, R. Rossignol, T. Letellier et al., “Hypoxia-inducible factor-1alpha, a key factor in the keratinocyte response to UVB exposure,” J. Biol. Chem. 282, 16413–16422 (2007). Web of Science, Google Scholar
41. L. I. Johnson-Cadwell, M. B. Jekabsons, A. Wang, B. M. Polster, D. G. Nicholls, “‘Mild Uncoupling’ does not decrease mitochondrial superoxide levels in cultured cerebellar granule neurons but decreases spare respiratory capacity and increases toxicity to glutamate and oxidative stress,” J. Neurochem. 101, 1619–1631 (2007). Web of Science, Google Scholar
42. B. A. Roelofs, S. X. Ge, P. E. Studlack, B. M. Polster, “Low micromolar concentrations of the superoxide probe MitoSOX uncouple neural mitochondria and inhibit complex IV,” Free Radic. Biol. Med. 86, 250–258 (2015). Web of Science, Google Scholar
43. B. Kalyanaraman, B. P. Dranka, M. Hardy, R. Michalski, J. Zielonka, “HPLC-based monitoring of products formed from hydroethidine-based fluorogenic probes - The ultimate approach for intra- and extracellular superoxide detection,” Biochim. Et Biophys. Acta-Gen. Subj. 1840, 739–744 (2014). Web of Science, Google Scholar
44. J. Zielonka, J. Vasquez-Vivar, B. Kalyanaraman, “Detection of 2-hydroxyethidium in cellular systems: A unique marker product of superoxide and hydroethidine,” Nat. Protoc. 3, 8–21 (2008). Web of Science, Google Scholar
45. H. J. Heusinkveld, R. H. S. Westerink, “Caveats and limitations of plate reader-based high-throughput kinetic measurements of intracellular calcium levels,” Toxicol. Appl. Pharmacol. 255, 1–8 (2011). Web of Science, Google Scholar
46. P. J. Bushway, M. Mercola, J. H. Price, “A comparative analysis of standard microtiter plate reading versus imaging in cellular assays,” Assay Drug Dev. Technol. 6, 557–567 (2008). Web of Science, Google Scholar
47. S. Menazza, E. Murphy, “The Expanding Complexity of Estrogen Receptor Signaling in the Cardiovascular System,” Circul. Res. 118, 994–1007 (2016). Web of Science, Google Scholar
48. X. Y. Li, P. Fang, J. T. Mai, E. T. Choi, H. Wang, X. F. Yang, “Targeting mitochondrial reactive oxygen species as novel therapy for inflammatory diseases and cancers,” J. Hematol. Oncol. 6, 6–19 (2013). Web of Science, Google Scholar
49. M. A. Aon, S. Cortassa, B. O’Rourke, “Redox-optimized ROS balance: A unifying hypothesis,” Biochim. Et Biophys. Acta-Bioenerg. 1797, 865–877 (2010). Web of Science, Google Scholar
50. C. Brueckl, S. Kaestle, A. Kerem, H. Habazettl, F. Krombach, H. Kuppe et al., “Hyperoxia-induced reactive oxygen species formation in pulmonary capillary endothelial cells in situ,” Am. J. Respir. Cell Mol. Biol. 34(4), 453–463 (2006). Web of Science, Google Scholar
51. J. F. Turrens, “Mitochondrial formation of reactive oxygen species,” J. Physiol. 552, 335–344 (2003). Web of Science, Google Scholar
52. R. H. Kallet, M. A. Matthay, “Hyperoxic acute lung injury,” Respir. Care 58, 123–141 (2013). Web of Science, Google Scholar
53. E. Cadenas, A. Boveris, C. I. Ragan, A. O. Stoppani, “Production of superoxide radicals and hydrogen peroxide by NADH-ubiquinone reductase and ubiquinol-cytochrome c reductase from beef-heart mitochondria,” Arch. Biochem. Biophys. 180, 248–257 (1977). Web of Science, Google Scholar
54. J. F. Turrens, A. Alexandre, A. L. Lehninger, “Ubisemiquinone is the electron donor for superoxide formation by complex III of heart mitochondria,” Arch. Biochem. Biophys. 237, 408–414 (1985). Web of Science, Google Scholar
55. Y. Liu, G. Fiskum, D. Schubert, “Generation of reactive oxygen species by the mitochondrial electron transport chain,” J. Neurochem. 80, 780–787 (2002). Web of Science, Google Scholar
56. L. Zhang, L. D. Yu, C. A. Yu, “Generation of superoxide anion by succinate-cytochrome c reductase from bovine heart mitochondria,” J. Biol. Chem. 273, 33972–33976 (1998). Web of Science, Google Scholar
57. P. R. Rich, W. D. Bonner, “The sites of superoxide anion generation in higher plant mitochondria,” Arch. Biochem. Biophys. 188, 206–213 (1978). Web of Science, Google Scholar
58. I. V. Grigolava, M. Ksenzenko, A. A. Konstantinob, A. N. Tikhonov, T. M. Kerimov, “[Tiron as a spin-trap for superoxide radicals produced by the respiratory chain of submitochondrial particles],” Biokhimiia 45, 75–82 (1980). Google Scholar
59. F. Diaz, H. Fukui, S. Garcia, C. T. Moraes, “Cytochrome c oxidase is required for the assembly/stability of respiratory complex I in mouse fibroblasts,” Mol. Cell. Biol. 26, 4872–4881 (2006). Web of Science, Google Scholar
60. H. B. Leavesley, L. Li, K. Prabhakaran, J. L. Borowitz, G. E. Isom, “Interaction of cyanide and nitric oxide with cytochrome c oxidase: Implications for acute cyanide toxicity,” Toxicol. Sci. 101, 101–111 (2008). Web of Science, Google Scholar
61. I. Sipos, L. Tretter, V. Adam-Vizi, “Quantitative relationship between inhibition of respiratory complexes and formation of reactive oxygen species in isolated nerve terminals,” J. Neurochem. 84, 112–118 (2003). Web of Science, Google Scholar
62. A. K. S. Camara, M. L. Riess, L. G. Kevin, E. Novalija, D. F. Stowe, “Hypothermia augments reactive oxygen species detected in the guinea pig isolated perfused heart,” Am. J. Physiol.-Heart Circul. Physiol. 286, H1289–H1299 (2004). Web of Science, Google Scholar
63. I. Amigo, F. M. da Cunha, M. F. Forni, W. Garcia-Neto, P. A. Kakimoto, L. A. Luevano-Martinez et al., “Mitochondrial form, function and signalling in aging,” Biochem. J. 473, 3421–3449 (2016). Web of Science, Google Scholar
64. P. G. Gunasekar, J. L. Borowitz, G. E. Isom, “Cyanide-induced generation of oxidative species: Involvement of nitric oxide synthase and cyclooxygenase-2,” J. Pharmacol. Exp. Therap. 285, 236–241 (998). Web of Science, Google Scholar
65. D. C. Jones, P. G. Gunasekar, J. L. Borowitz, G. E. Isom, “Dopamine-induced apoptosis is mediated by oxidative stress and is enhanced by cyanide in differentiated PC12 cells,” J. Neurochem. 74, 2296–2304 (2000). Web of Science, Google Scholar
66. Q. Chen, E. J. Vazquez, S. Moghaddas, C. L. Hoppel, E. J. Lesnefsky, “Production of reactive oxygen species by mitochondria - Central role of complex III,” J. Biolog. Chem. 278, 36027–36031 (2003). Web of Science, Google Scholar
67. Y. J. Wang, Y. S. Ho, S. W. Chu, H. J. Lien, T. H. Liu, J. K. Lin, “Induction of glutathione depletion, p53 protein accumulation and cellular transformation by tetrachlorohydroquinone, a toxic metabolite of pentachlorophenol (vol 105, pg 1, 1997),” Chem.-Biolog. Interac. 106, 1–16 (1997). Web of Science, Google Scholar
68. W. C. Dorsey, P. B. Tchounwou, B. D. Ford, “Neuregulin 1-Beta cytoprotective role in AML 12 mouse hepatocytes exposed to pentachlorophenol,” Int. J. Environ. Res. Publ. Health 3, 11–22 (2006). Google Scholar
69. P. Fernandez Freire, V. Labrador, J. M. P. Martin, M. J. Hazen, “Cytotoxic effects in mammalian Vero cells exposed to pentachlorophenol,” Toxicology 210, 37–44 (2005). Web of Science, Google Scholar
70. J. Folch, M. Yeste-Velasco, D. Alvira, A. V. de la Torre, M. Bordas, M. Lopez et al., “Evaluation of pathways involved in pentachlorophenol-induced apoptosis in rat neurons,” Neurotoxicology 30, 451–458 (2009). Web of Science, Google Scholar
71. R. Sepehr, S. H. Audi, K. S. Staniszewski, S. T. Haworth, E. R. Jacobs, M. Ranji, “Novel Flurometric Tool to Assess Mitochondrial Redox State of Isolated Perfused Rat Lungs after Exposure to Hyperoxia,” IEEE J. Transl. Eng. Health. Med. 1, (2013). Google Scholar
72. Y. L. Dong, P. J. Zhou, S. Y. Jiang, X. W. Pan, X. H. Zhao, “Induction of oxidative stress and apoptosis by pentachlorophenol in primary cultures of Carassius carassius hepatocytes,” Comparative Biochem. Physiol. C-Toxicol. Pharmacol. 150, 179–185 (2009). Web of Science, Google Scholar
73. J. N. He, Y. Duan, D. P. Hua, G. J. Fan, L. Wang, Y. Liu et al., “DEXH Box RNA Helicase-Mediated Mitochondrial Reactive Oxygen Species Production in Arabidopsis Mediates Crosstalk between Abscisic Acid and Auxin Signaling,” Plant Cell 24, 1815–1833 (2012). Web of Science, Google Scholar
74. R. F. Feissner, J. Skalska, W. E. Gaum, S. S. Sheu, “Crosstalk signaling between mitochondrial Ca2 $+$ and ROS,” Front. Biosci.-Landmark 14, 1197–1218 (2009). Web of Science, Google Scholar
75. E. Cadenas, A. Boveris, “Enhancement of hydrogen peroxide formation by protophores and ionophores in antimycin-supplemented mitochondria,” Biochem. J. 188, 31–37 (1980). Web of Science, Google Scholar
76. B. D. Fink, Y. O’Malley, B. L. Dake, N. C. Ross, T. E. Prisinzano, W. I. Sivitz, “Mitochondrial targeted coenzyme Q, superoxide, and fuel selectivity in endothelial cells,” PLoS One 4, e4250 (2009). Web of Science, Google Scholar
77. M. F. Ross, G. F. Kelso, F. H. Blaikie, A. M. James, H. M. Cocheme, A. Filipovska et al., “Lipophilic triphenylphosphonium cations as tools in mitochondrial bioenergetics and free radical biology,” Biochem.-Moscow 70, 222–230 (2005). Web of Science, Google Scholar
78. M. Kalbacova, M. Vrbacky, Z. Drahota, Z. Melkova, “Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry,” Cytometry A 52, 110–116 (2003). Web of Science, Google Scholar
79. A. R. Khaled, D. A. Reynolds, H. A. Young, C. B. Thompson, K. Muegge, S. K. Durum, “Interleukin-3 withdrawal induces an early increase in mitochondrial membrane potential unrelated to the Bcl-2 fammily - Roles of intracellular pH, ADP transport, and F0F1-ATPase,” J. Biolog. Chem. 276, 6453–6462 (2001). Web of Science, Google Scholar
80. F. M. P. de Gannes, M. A. Belaud-Rotureau, P. Voisin, N. Leducq, F. Belloc, P. Canioni et al., “Flow cytometric analysis of mitochondrial activity in situ: Application to acetylceramide-induced mitochondrial swelling and apoptosis,” Cytometry 33, 333–339 (1998). Google Scholar

Vol. 11, No. 01

Metrics

Downloaded 1,022 times

History

Received 9 February 2017

Accepted 1 May 2017

Published: 30 June 2017

Information

This is an Open Access article published by World Scientific Publishing Company. It is distributed under the terms of the Creative Commons Attribution 4.0 (CC-BY) License. Further distribution of this work is permitted, provided the original work is properly cited.

Keywords

PDF download

System Upgrade on Tue, May 28th, 2024 at 2am (EDT)

Quantitative optical measurement of mitochondrial superoxide dynamics in pulmonary artery endothelial cells

Abstract

Recommended