Song 2024 Commun Biol
| Song A, Zhao N, Hilpert DC, Perry C, Baur JA, Wallace DC, Schaefer PM (2024) Visualizing subcellular changes in the NAD(H) pool size versus redox state using fluorescence lifetime imaging microscopy of NADH. Commun Biol 7:428. https://doi.org/10.1038/s42003-024-06123-7 |
Song Angela, Zhao Nicole, Hilpert Diana C, Perry Caroline, Baur Joseph A, Wallace Douglas C, Schaefer Patrick M (2024) Commun Biol
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.
• Bioblast editor: Plangger M • O2k-Network Lab: US PA Philadelphia Baur JA, US PA Philadelphia Wallace DC, DE Ulm Radermacher P
Labels: MiParea: Respiration
Organism: Human
Tissue;cell: HEK
Preparation: Intact cells
Coupling state: LEAK, ROUTINE, ET
Pathway: ROX
HRR: Oxygraph-2k
2024-04
