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.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“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
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Principles of Fluorescence Spectroscopy
Lakowicz, J. R. (1999) · Springer eBooks · doi:10.1007/978-1-4757-3061-6 · Cited by 26,948
Understanding the Warburg Effect: The Metabolic Requirements of Cell Proliferation
Vander Heiden, M. G., Cantley, L. C., & Thompson, C. B. (2009) · Science · doi:10.1126/science.1160809 · Cited by 16,197
Warburg, O. (1956) · Science · doi:10.1126/science.123.3191.309 · Cited by 13,425
Shaner, N. C., Campbell, R. E., Steinbach, P. A., Giepmans, B. N. G., Palmer, A. E., & Tsien, R. Y. (2004) · Nature Biotechnology · doi:10.1038/nbt1037 · Cited by 4,619
Pellerin, L., & Magistretti, P. J. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.22.10625 · Cited by 2,927 · cited 1×
“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).”
Debnath, J., Muthuswamy, S. K., & Brugge, J. S. (2003) · Methods · doi:10.1016/s1046-2023(03)00032-x · Cited by 2,001
Williamson, D., Lund, P., & Krebs, H. (1967) · Biochemical Journal · doi:10.1042/bj1030514 · Cited by 1,871
Genetically encoded fluorescent indicator for intracellular hydrogen peroxide
Belousov, V. V., Fradkov, A. F., Lukyanov, K. A., Staroverov, D. B., Shakhbazov, K., Terskikh, A. V., & Lukyanov, S. (2006) · Nature Methods · doi:10.1038/nmeth866 · Cited by 1,286
Regulation of the mTOR Complex 1 Pathway by Nutrients, Growth Factors, and Stress
Sengupta, S., Peterson, T. R., & Sabatini, D. M. (2010) · Molecular Cell · doi:10.1016/j.molcel.2010.09.026 · Cited by 1,275
Short-chain dehydrogenases/reductases (SDR)
Jörnvall, H., Persson, B., Krook, M., Atrian, S., Gonzàlez‐Duarte, R., Jeffery, J., & Ghosh, D. (1995) · Biochemistry · doi:10.1021/bi00018a001 · Cited by 1,241
Maira, S.-M., Stauffer, F., Brueggen, J., Furet, P., Schnell, C., Fritsch, C., Brachmann, S., Chène, P., De Pover, A., Schoemaker, K., et al. (2008) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-08-0017 · Cited by 1,134
Reducing the Environmental Sensitivity of Yellow Fluorescent Protein
Griesbeck, O., Baird, G. S., Campbell, R. E., Zacharias, D. A., & Tsien, R. Y. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m102815200 · Cited by 1,023
Bitterman, K. J., Anderson, R. M., Cohen, H. Y., Latorre-Esteves, M., & Sinclair, D. A. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m205670200 · Cited by 1,002
Circularly permuted green fluorescent proteins engineered to sense Ca 2+
Nagai, T., Sakaue-Sawano, A., Park, E. S., & Miyawaki, A. (2001) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.051636098 · Cited by 969
Circular permutation and receptor insertion within green fluorescent proteins
Baird, G. S., Zacharias, D. A., & Tsien, R. Y. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.20.11241 · Cited by 963
Intracellular Oxidation-Reduction States in Vivo
Chance, B., Cohen, P., Jöbsis, F. F., & Schoener, B. (1962) · Science · doi:10.1126/science.137.3529.499 · Cited by 947
Cytosolic phosphorylation potential.
Veech, R. L., Lawson, J., Cornell, N. W., & Krebs, H. A. (1979) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)50401-4 · Cited by 919
Nampt/PBEF/Visfatin Regulates Insulin Secretion in β Cells as a Systemic NAD Biosynthetic Enzyme
Revollo, J. R., Körner, A., Mills, K. F., Satoh, A., Wang, T., Garten, A., Dasgupta, B., Sasaki, Y., Wolberger, C., Townsend, R. R., et al. (2007) · Cell Metabolism · doi:10.1016/j.cmet.2007.09.003 · Cited by 911
Herrero-Mendez, A., Almeida, A., Fernández, E., Maestre, C., Moncada, S., & Bolaños, J. P. (2009) · Nature Cell Biology · doi:10.1038/ncb1881 · Cited by 844
Brain metabolism dictates the polarity of astrocyte control over arterioles
Gordon, G. R., Choi, H. B., Rungta, R. L., Ellis‐Davies, G. C. R., & MacVicar, B. A. (2008) · Nature · doi:10.1038/nature07525 · Cited by 770
Neural activity triggers neuronal oxidative metabolism followed by astrocytic glycolysis
Kasischke, K. A., Vishwasrao, H. D., Fisher, P. J., Zipfel, W. R., & Webb, W. W. (2004) · Science · doi:10.1126/science.1096485 · Cited by 742
Determination of the absolute quantum yield of fluorescent solutions
Weber, G., & Teale, F. W. J. (1957) · Transactions of the Faraday Society · doi:10.1039/tf9575300646 · Cited by 665
Regulation of Corepressor Function by Nuclear NADH
Zhang, Q., Piston, D. W., & Goodman, R. H. (2002) · Science · doi:10.1126/science.1069300 · Cited by 620
CellProfiler Analyst: data exploration and analysis software for complex image-based screens
Jones, T. R., Kang, I. H., Wheeler, D. B., Lindquist, R. A., Papallo, A., Sabatini, D. M., Golland, P., & Carpenter, A. E. (2008) · BMC Bioinformatics · doi:10.1186/1471-2105-9-482 · Cited by 543
THE FLUOROMETRIC MEASUREMENT OF PYRIDINE NUCLEOTIDES
Lowry, O. H., Roberts, N. R., & Kapphahn, J. I. (1957) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)64996-8 · Cited by 532
A genetically encoded fluorescent reporter of ATP:ADP ratio
Berg, J., Hung, Y. P., & Yellen, G. (2009) · Nature Methods · doi:10.1038/nmeth.1288 · Cited by 493
Eto, K., Tsubamoto, Y., Terauchi, Y., 杉山, 拓., Kishimoto, T. E., Takahashi, N. K., Yamauchi, N., Kubota, N., Murayama, S., Aizawa, T., et al. (1999) · Science · doi:10.1126/science.283.5404.981 · Cited by 492
Garriga-Canut, M., Schoenike, B. A., Qazi, R., Bergendahl, K., Daley, T. J., Pfender, R. M., Morrison, J. F., Ockuly, J. C., Stafstrom, C. E., Sutula, T. P., et al. (2006) · Nature Neuroscience · doi:10.1038/nn1791 · Cited by 446
Imaging Intracellular pH in Live Cells with a Genetically Encoded Red Fluorescent Protein Sensor
Tantama, M., Hung, Y. P., & Yellen, G. (2011) · Journal of the American Chemical Society · doi:10.1021/ja202902d · Cited by 411
Scott, T. G., Spencer, R. D., Leonard, N. J., & Weber, G. (1970) · Journal of the American Chemical Society · doi:10.1021/ja00706a043 · Cited by 389
Perraud, A.-L., Takanishi, C. L., Shen, B. W., Kang, S. J., Smith, M., Schmitz, C., Knowles, H. M., Ferraris, D. V., Li, W., Zhang, J., et al. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m411446200 · Cited by 362
Bartlett, W., & Banker, G. (1984) · Journal of Neuroscience · doi:10.1523/jneurosci.04-08-01944.1984 · Cited by 348
Akerboom, J., Rivera, J. D. V., Rodríguez‐Guilbe, M. M., Malavé, E. C. A., Hernández, H. H., Tian, L., Hires, S. A., Marvin, J. S., Looger, L. L., & Schreiter, E. R. (2008) · Journal of Biological Chemistry · doi:10.1074/jbc.m807657200 · Cited by 302
Transcription Corepressor CtBP Is an NAD+-Regulated Dehydrogenase
Kumar, V., Carlson, J. E., Ohgi, K. A., Edwards, T. A., Rose, D. W., Escalante, C. R., Rosenfeld, M. G., & Aggarwal, A. K. (2002) · Molecular Cell · doi:10.1016/s1097-2765(02)00650-0 · Cited by 299
Patterson, G. H., Knobel, S. M., Arkhammar, P. O. G., Thastrup, M., & Piston, D. W. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.090098797 · Cited by 284
Regulation of redox metabolism in the mouse oocyte and embryo
Dumollard, R., Ward, Z., Carroll, J., & Duchen, M. R. (2006) · Development · doi:10.1242/dev.02744 · Cited by 246
A novel sensor of NADH/NAD+ redox poise in Streptomyces coelicolor A3(2)
Brekasis, D., & Paget, M. S. B. (2003) · The EMBO Journal · doi:10.1093/emboj/cdg453 · Cited by 224
GFP-like Proteins Stably Accumulate in Lysosomes
Katayama, H., Yamamoto, A., Mizushima, N., Yoshimori, T., & Miyawaki, A. (2008) · Cell Structure and Function · doi:10.1247/csf.07011 · Cited by 223
Redox sensor CtBP mediates hypoxia-induced tumor cell migration
Zhang, Q., Wang, S.-Y., Nottke, A. C., Rocheleau, J. V., Piston, D. W., & Goodman, R. H. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0603269103 · Cited by 200
(1990) · Elsevier eBooks · doi:10.1016/b978-0-12-584552-6.x5001-0 · Cited by 195
Rocheleau, J. V., Head, W. S., & Piston, D. W. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m314005200 · Cited by 195
Efficiently folding and circularly permuted variants of the Sapphire mutant of GFP
Zapata-Hommer, O., & Griesbeck, O. (2003) · BMC Biotechnology · doi:10.1186/1472-6750-3-5 · Cited by 170
Bücher, T., Brauser, B., CONZE, A., KLEIN, F., Langguth, O., & Sies, H. (1972) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1972.tb01840.x · Cited by 165
Nausch, L., Ledoux, J., Bonev, A. D., Nelson, M. T., & Dostmann, W. R. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0710387105 · Cited by 164
NAD(P)H fluorescence imaging of postsynaptic neuronal activation in murine hippocampal slices
Shuttleworth, C. W., Brennan, A. M., & Connor, J. A. (2003) · The Journal of Neuroscience · doi:10.1523/jneurosci.23-08-03196.2003 · Cited by 151
The redox switch/redox coupling hypothesis
Cerdán, S., Rodrigues, T. B., Sierra, A., Benito, M., Fonseca, L. L., Fonseca, C. P., & García-Martín, M. L. (2006) · Neurochemistry International · doi:10.1016/j.neuint.2005.12.036 · Cited by 146
Structure and functional properties of the Bacillus subtilis transcriptional repressor Rex
Wang, E., Bauer, M. C., Rogstam, A., Linse, S. S., Logan, D. T., & von Wachenfeldt, C. (2008) · Molecular Microbiology · doi:10.1111/j.1365-2958.2008.06295.x · Cited by 143
Klingenberg, M., & Bücher, T. (1960) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.29.070160.003321 · Cited by 137
Fluorescence of Pyridine Nucleotides in Mitochondria
Avi‐Dor, Y., Olson, J. M., Doherty, M. D., & Kaplan, N. O. (1962) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)63448-4 · Cited by 127
Glucose-induced changes in cytosolic ATP content in pancreatic islets
Malaisse, W. J., & Sener, A. (1987) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/0167-4889(87)90134-0 · Cited by 116 · cited 1×
“Although metabolic challenges often lead to energy deprivation and decrease in cytosolic ATP:ADP ratio, changes in cytosolic adenine nucleotide pool size are minimal (Schwenke et al., 1981; Malaisse and Sener, 1987) and would not be expected to interfere with Peredox response.”
Mitochondrial and cytosolic ATP/ADP ratios in rat liver in vivo
Schwenke, W. D., Soboll, S., Seitz, H. J., & Sies, H. (1981) · Biochemical Journal · doi:10.1042/bj2000405 · Cited by 114
Intracellular pH Sensing Using Autofluorescence Lifetime Microscopy
Ogikubo, S., Nakabayashi, T., Adachi, T., Islam, M. S., Yoshizawa, T., Kinjo, M., & Ohta, N. (2011) · The Journal of Physical Chemistry B · doi:10.1021/jp2058904 · Cited by 99
Structural Basis for NADH/NAD+ Redox Sensing by a Rex Family Repressor
McLaughlin, K. J., Strain-Damerell, C. M., Xie, K., Brekasis, D., Soares, A. S., Paget, M. S. B., & Kielkopf, C. L. (2010) · Molecular Cell · doi:10.1016/j.molcel.2010.05.006 · Cited by 93
X-Ray Structure of a Rex-Family Repressor/NADH Complex Insights into the Mechanism of Redox Sensing
Sickmier, E. A., Brekasis, D., Paranawithana, S. R., Bonanno, J. B., Paget, M. S. B., Burley, S. K., & Kielkopf, C. L. (2005) · Structure · doi:10.1016/j.str.2004.10.012 · Cited by 87
PROPERTIES OF THE COELENTERATE GREEN-FLUORESCENT PROTEINS
Ward, W. W. (1981) · Elsevier eBooks · doi:10.1016/b978-0-12-208820-9.50035-5 · Cited by 71
The conserved NAD(H)-dependent corepressor CTBP-1 regulates Caenorhabditis elegans life span
Chen, S., Whetstine, J. R., Ghosh, S. K., Hanover, J. A., Gali, R. R., Grosu, P., & Shi, Y. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0802674106 · Cited by 67
Oxygen partial pressure and free intracellular adenosine of isolated cardiomyocytes
Smolenski, R. T., Schrader, J., de Groot, H., & Deussen, A. (1991) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.1991.260.4.c708 · Cited by 50
Adenine nucleotides and the adenylate kinase equilibrium in livers of foetal and newborn rats
Ballard, F. J. (1970) · Biochemical Journal · doi:10.1042/bj1170231 · Cited by 39
Flagged first, then by citations
Cited by
Sorted by citations
Understanding the Intersections between Metabolism and Cancer Biology
Heiden, M. G. V., & DeBerardinis, R. J. (2017) · Cell · doi:10.1016/j.cell.2016.12.039 · Cited by 2,369
Fueling Immunity: Insights into Metabolism and Lymphocyte Function
Pearce, E. L., Poffenberger, M. C., Chang, C.-H., & Jones, R. G. (2013) · Science · doi:10.1126/science.1242454 · Cited by 1,363
Supporting aspartate biosynthesis is an essential function of respiration in proliferating cells
Sullivan, L. B., Gui, D. Y., Hosios, A. M., Bush, L. N., Freinkman, E., & Vander Heiden, M. G. (2015) · Cell · doi:10.1016/j.cell.2015.07.017 · Cited by 1,173
NAD(H) and NADP(H) Redox Couples and Cellular Energy Metabolism
Xiao, W., Wang, R., Handy, D. E., & Loscalzo, J. (2017) · Antioxidants and Redox Signaling · doi:10.1089/ars.2017.7216 · Cited by 1,062
Brain Glucose Metabolism: Integration of Energetics with Function
Dienel, G. A. (2018) · Physiological Reviews · doi:10.1152/physrev.00062.2017 · Cited by 911
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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
