The DOI matches its Crossref record; 58 of the 58 references it cites were found in a registry or an index.
Imaging cytosolic NADH-NAD(+) redox state with a genetically encoded fluorescent biosensor
Open accessAbstract
NADH is a key metabolic cofactor whose sensitive and specific detection in the cytosol of live cells has been difficult. We constructed a fluorescent biosensor of the cytosolic NADH-NAD(+) redox state by combining a circularly permuted GFP T-Sapphire with a bacterial NADH-binding protein, Rex. Although the initial construct reported [NADH] × [H(+)] / [NAD(+)], its pH sensitivity was eliminated by mutagenesis. The engineered biosensor Peredox reports cytosolic NADH:NAD(+) ratios and can be calibrated with exogenous lactate and pyruvate. We demonstrated its utility in several cultured and primary cell types. We found that glycolysis opposed the lactate dehydrogenase equilibrium to produce a reduced cytosolic NADH-NAD(+) redox state. We also observed different redox states in primary mouse astrocytes and neurons, consistent with hypothesized metabolic differences. Furthermore, using high-content image analysis, we monitored NADH responses to PI3K pathway inhibition in hundreds of live cells. As an NADH reporter, Peredox should enable better understanding of bioenergetics.
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“However, the use of pure lactate as an alternative fuel, rather than a redox-balanced mixture of lactate and pyruvate, abnormally elevates the low cytosolic NADH redox ratio in neurons (seen in vitro by Hung et al. [2011] and Mongeon et al. [2016] as well as in vivo by Díaz-García et al. [2017]); this decreases the rate of neuronal glycolysis in a way that would not represent the normal effect in”
From Fueling thought: Management of glycolysis and oxidative phosphorylation in neuronal metabolism (2018) · cites this paper as Hung et al., 2011
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“Primary Cultured Cortical Astrocytes and Neurons Differ in Their Cytosolic NADH-NAD Redox States Glucose metabolism has been proposed to differ in astrocytes and neurons (Pellerin and Magistretti, 1994; Herrero-Mendez et al., 2009).”
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Metadata
- Authors
- Hung, Y. P., Albeck, J. G., Tantama, M., & Yellen, G.
- Affiliations
- Harvard University (US)
- Corresponding author
- Yellen, Gary
- Venue
- Cell Metabolism
- Publisher
- Elsevier BV
- Published
- 2011-10
- Volume
- 14
- Issue
- 4
- Pages
- 545-554
- ISSN
- 1550-4131 (ISSN-L)
- PMC
- PMC3190165
- OpenAlex
- W2155184340
- Cited by
- 509
- References
- 58
- Citations by year
- 2026: 31 · 2025: 29 · 2024: 32 · 2023: 34 · 2022: 34 · 2021: 43 · 2020: 59 · 2019: 29 · 2018: 46 · 2017: 39 · 2016: 30 · 2015: 29 · 2014: 34 · 2013: 26 · 2012: 12
- FWCI
- 13.40
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 655 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Cell Metabolism (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- U.S. Army (W81XWH-08-1-0609)
- National Institutes of Health (R01 NS055031, F32 NS066613)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
- Keywords
- genetically encoded fluorescent sensors, circularly permuted GFP, glycolysis, lactate dehydrogenase, astrocytes, neurons, bioenergetics, live-cell imaging
- MeSH terms
- Phosphoinositide-3 Kinase Inhibitors, Animals, Bacterial Proteins, Cells, Cultured, Cytosol, Glycolysis, Hydrogen-Ion Concentration, L-Lactate Dehydrogenase, Luminescent Proteins, NAD, Oxidation-Reduction, Recombinant Fusion Proteins, Biosensing Techniques, Phosphatidylinositol 3-Kinases, Mice
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