Journal
NATURE COMMUNICATIONS
Volume 5, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/ncomms4936
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Funding
- Action on Hearing Loss
- Wellcome Trust-UCL Therapeutic Innovation Fund, Telethon (Italy) [GEP1206]
- Italian Association for Cancer Research (AIRC)
- Wellcome Trust/MRC Joint Call in Neurodegeneration Award [WT089698]
- CoMPLEX Doctoral Training Centre at UCL
- Parkinson's UK
- British Heart Foundation
- BBSRC [BB/L020874/1] Funding Source: UKRI
- MRC [MR/K000608/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/L020874/1] Funding Source: researchfish
- Medical Research Council [MR/K000608/1] Funding Source: researchfish
- RNID [G52] Funding Source: researchfish
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NAD is a key determinant of cellular energy metabolism. In contrast, its phosphorylated form, NADP, plays a central role in biosynthetic pathways and antioxidant defence. The reduced forms of both pyridine nucleotides are fluorescent in living cells but they cannot be distinguished, as they are spectrally identical. Here, using genetic and pharmacological approaches to perturb NAD(P)H metabolism, we find that fluorescence lifetime imaging (FLIM) differentiates quantitatively between the two cofactors. Systematic manipulations to change the balance between oxidative and glycolytic metabolism suggest that these states do not directly impact NAD(P)H fluorescence decay rates. The lifetime changes observed in cancers thus likely reflect shifts in the NADPH/NADH balance. Using a mathematical model, we use these experimental data to quantify the relative levels of NADH and NADPH in different cell types of a complex tissue, the mammalian cochlea. This reveals NADPH-enriched populations of cells, raising questions about their distinct metabolic roles.
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