Faculty Collaboration Database

Surface fluorescence studies of tissue mitochondrial redox state in isolated perfused rat lungs. Ann Biomed Eng 2013 Apr;41(4):827-36

Date

12/15/2012

Pubmed ID

23238793

Pubmed Central ID

PMC3606690

DOI

10.1007/s10439-012-0716-z

Scopus ID

2-s2.0-84876466897 (requires institutional sign-in at Scopus site)   20 Citations

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.

Author List

Staniszewski K, Audi SH, Sepehr R, Jacobs ER, Ranji M

Authors

Said Audi PhD Professor in the Biomedical Engineering department at Marquette University
Mahsa Ranji PhD Assistant Professor in the Electrical Engineering department at University of Wisconsin - Milwaukee

MESH terms used to index this publication - Major topics in bold

Animals
Biomedical Engineering
Equipment Design
Flavin-Adenine Dinucleotide
Fluorometry
In Vitro Techniques
Lung
Male
Mitochondria
NAD
Optical Fibers
Oxidation-Reduction
Perfusion
Rats
Rats, Sprague-Dawley