Assessment of mitochondrial metabolic oxidative state in living cardiomyocytes with spectrally-resolved fluorescence lifetime spectroscopy of NAD(P)H

• Published in: Sheng wu yi xue gong cheng xue za zhi Vol. 26; no. 6; p. 1191
• Main Authors: Cheng, Ying, Ren, Mingming, Niu, Yanyan, Qiao, Jianhua, Aneba, S, Chorvat, Jr, D, Chorvatova, A
• Format: Journal Article
• Language: Chinese
• Published: China 01.12.2009
• Subjects: Animals; In Vitro Techniques; Mitochondria, Heart - metabolism; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; NADP - analysis; NADP - metabolism; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Spectrometry, Fluorescence - methods
• ISSN: 1001-5515

The primary function of cardiac mitochondria is the production of ATP to support heart contraction. Examination of the mitochondrial redox state is therefore crucially important to sensitively detect early signs of mitochondrial function in pathophysiological conditions, such as ischemia, diabetes and heart failure. We study fingerprinting of mitochondrial metabolic oxidative state in living cardiomyocytes with spectrally-resolved fluorescence lifetime spectroscopy of NAD(P)H, the principal electron donor in mitochondrial respiration responsible for vital ATP supply. Here NAD(P)H is studied as a marker for non-invasive fluorescent probing of the mitochondrial function. NAD(P) H fluorescence is recorded in cardiac cells following excitation with 375nm UV-light and detection by spectrally-resolved time-correlated single photon counting (TCSPC), based on the simultaneous measurement of the fluorescence spectra and fluorescence lifetimes. Modulation of NADH production and/or mitochondrial respiration is tested to