Separating NADH and NADPH fluorescence in live cells and tissues using FLIM
University College London · University of Bergen · +2 more institutions
Abstract
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…
Citation impact
- FWCI
- 52.60
- Percentile
- 100%
- References
- 58
Authors
7Topics & keywords
- NAD+ kinase
- Fluorescence
- Oxidative phosphorylation
- Cofactor
- Biochemistry
- Biology
- Metabolism
- Fluorescence-lifetime imaging microscopy
- Affordable and clean energy
Funding
- WTWellcome TrustAward: WT089698
- BHBritish Heart Foundation
- PUParkinson's UK
- AIAssociazione Italiana per la Ricerca sul Cancro
- MRMedical Research CouncilAwards: WT089698, MR/K000608/1
- EAEngineering and Physical Sciences Research Council
- BABiotechnology and Biological Sciences Research CouncilAward: BB/L020874/1