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Annals of Biomedical Engineering

, Volume 41, Issue 4, pp 827–836 | Cite as

Surface Fluorescence Studies of Tissue Mitochondrial Redox State in Isolated Perfused Rat Lungs

  • Kevin Staniszewski
  • Said H. Audi
  • Reyhaneh Sepehr
  • Elizabeth R. Jacobs
  • Mahsa RanjiEmail author
Article

Abstract

We designed a fiber-optic-based optoelectronic fluorometer to measure emitted fluorescence from the auto-fluorescent electron carriers NADH and FAD of the mitochondrial electron transport chain (ETC). The ratio of NADH to FAD is called the redox ratio (RR = NADH/FAD) and is an indicator of the oxidoreductive state of tissue. We evaluated the fluorometer by measuring the fluorescence intensities of NADH and FAD at the surface of isolated, perfused rat lungs. Alterations of lung mitochondrial metabolic state were achieved by the addition of rotenone (complex I inhibitor), potassium cyanide (KCN, complex IV inhibitor) and/or pentachlorophenol (PCP, uncoupler) into the perfusate recirculating through the lung. Rotenone- or KCN-containing perfusate increased RR by 21 and 30%, respectively. In contrast, PCP-containing perfusate decreased RR by 27%. These changes are consistent with the established effects of rotenone, KCN, and PCP on the redox status of the ETC. Addition of blood to perfusate quenched NADH and FAD signal, but had no effect on RR. This study demonstrates the capacity of fluorometry to detect a change in mitochondrial redox state in isolated perfused lungs, and suggests the potential of fluorometry for use in in vivo experiments to extract a sensitive measure of lung tissue health in real-time.

Keywords

Lung surface fluorometry Nicotinamide Adenine Dinucleotide (NADH) Flavin Adenine Dinucleotide (FADH2) Mitochondrial redox 

Notes

Acknowledgments

We appreciate the support of University of Wisconsin Milwaukee RGI 6 Grant, Clinical and Translational Science Institute KL2 Grant: NIH 8Kl2TR000056, Wisconsin Applied Research grant (Wi-ARG), NIH Grants HL-24349 (SHA), HL 49294 (ERJ), and the Department of Veterans Affairs that provided resources essential to the completion of these investigations. We acknowledge the technical help from Dr. Steven Haworth in the Clement J. Zablocki VA, Milwaukee, WI.

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Copyright information

© Biomedical Engineering Society 2012

Authors and Affiliations

  • Kevin Staniszewski
    • 1
  • Said H. Audi
    • 2
  • Reyhaneh Sepehr
    • 1
  • Elizabeth R. Jacobs
    • 3
    • 4
  • Mahsa Ranji
    • 1
    Email author
  1. 1.Biophotonics Lab, Department of Electrical EngineeringUniversity of Wisconsin MilwaukeeMilwaukeeUSA
  2. 2.Department of Biomedical EngineeringMarquette UniversityMilwaukeeUSA
  3. 3.Research and DevelopmentClement J. Zablocki VA Medical CenterMilwaukeeUSA
  4. 4.Medical College of WisconsinMilwaukeeUSA

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