Visualizing subcellular changes in the NAD(H) pool size versus redox state using fluorescence lifetime imaging microscopy of NADH
Year: 2024
Type: article
Abstract: NADH autofluorescence imaging is a promising approach for visualizing energy metabolism at the single-cell level. However, it is sensitive to the redox ratio and the total NAD(H) amount, which can change independently from each other, for example with aging. Here, we evaluate the potential of fluorescence lifetime imaging microscopy (FLIM) of NADH to differentiate between these modalities.We perform targeted modifications of the NAD(H) pool size and ratio in cells and mice and assess the impact on NADH FLIM. We show that NADH FLIM is sensitive to NAD(H) pool size, mimicking the effect of redox alterations. However, individual components of the fluorescence lifetime are differently impacted by redox versus pool size changes, allowing us to distinguish both modalities using only FLIM. Our results emphasize NADH FLIM's potential for evaluating cellular metabolism and relative NAD(H) levels with high spatial resolution, providing a crucial tool for our understanding of aging and metabolism.
Source: Communications Biology
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,890
Funders: U.S. Department of Defense, U.S. Department of Health and Human Services, University of Pennsylvania, Deutsche Forschungsgemeinschaft, National Institutes of Health, National Heart, Lung, and Blood Institute, National Institute of Mental Health, National Institute of Neurological Disorders and Stroke
Awards: G1433873921, G2262409748, G23045533, G3310788078, G3979633579, G4119534169, G4205532640, A MITOCHONDRIAL-INTERNEURONAL HYPOTHESIS OF AUTISM, COVID-19 SARS-COV-2 PROPAGATION AND PATHOLOGY AND MITOCHONDRIAL GENETICS, G5407178560, G5850682332, G6502886799, Mitochondrial Inborn Errors in Metabolism, Mouse models for mitochondrial disorders caused by mutations in mtDNA, G7003707359, POWERING UP MITOCHONDRIAL FUNCTIONS TO TREAT MITOCHONDRIAL DISEASE, G8390724461, Understanding the roles of cardiac NAD pools and therapeutic effects of precursor supplements in heart failure
OpenAlex ID: W4394604183