Measurement of glucose uptake and intracellular calcium concentration in single, living pancreatic beta-cells.
AuthorsYamada K, Nakata M, Horimoto N, Saito M, Matsuoka H, Inagaki N
JournalJ Biol Chem
PubMed ID10748091
'There has been no method previously to measure both glucose transport and its effect on the various intracellular functions in single, living mammalian cells. A fluorescent derivative of d-glucose, 2-[N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG), that we have developed has made such measurements possible. COS-1 cells that overexpress the human glucose transporter GLUT2 ... More
Glutamate triggers rapid glucose transport stimulation in astrocytes as evidenced by real-time confocal microscopy.
AuthorsLoaiza A, Porras OH, Barros LF
JournalJ Neurosci
PubMed ID12917367
'Glutamate stimulates glycolysis in astrocytes, a phenomenon that couples astrocytic metabolism with neuronal activity. However, it is not known whether glutamate also affects glucose transporter-1 (GLUT1), the transporter responsible for glucose entry into astrocytes. To address this question, two different real-time single-cell hexose uptake assays were applied to cultured hippocampal ... More
Fluorescent microplate cell assay to measure uptake and metabolism of glucose in normal human lung fibroblasts.
'This is the first report of a fluorimetric microplate assay to assess glucose uptake and metabolism in eukaryotic cells. The assay was carried out incubating normal human lung fibroblasts in the wells of microtiter trays with a fluorescent D-glucose derivative, 2-N-7-(nitrobenz-2-oxa-1,3-diazol-4-yl)amino-2-deoxy-D-glucose (2-NBDG). This dye could be incorporated by glucose transporting ... More
Rapid viability assessment of yeast cells using vital staining with 2-NBDG, a fluorescent derivative of glucose.
AuthorsOh KB, Matsuoka H
JournalInt J Food Microbiol
PubMed ID12038577
'A fluorescent glucose analogue, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG), which had been developed previously for the analysis of glucose uptake activity by living cells, was investigated to evaluate its applicability for assaying the viability of yeasts. Fluorescence intensities of the yeast population were measured by fluorescence spectrophotometry upon exposure to antifungal agents ... More
Intracellular fate of 2-NBDG, a fluorescent probe for glucose uptake activity, in Escherichia coli cells.
AuthorsYoshioka K, Saito M, Oh KB, Nemoto Y, Matsuoka H, Natsume M, Abe H
JournalBiosci Biotechnol Biochem
PubMed ID8987871
'A fluorescent derivative of D-glucose, 2-NBDG, which was previously developed for the evaluation of glucose uptake activity by living cells, was used on Escherichia coli cells and its fate after incorporation in the cells was investigated. 2-NBDG was converted to another fluorescent derivative (2-NBDG metabolite) immediately after it was taken ... More
p53 regulates glucose metabolism through an IKK-NF-kappaB pathway and inhibits cell transformation.
AuthorsKawauchi K, Araki K, Tobiume K, Tanaka N,
JournalNat Cell Biol
PubMed ID18391940
'Cancer cells use aerobic glycolysis preferentially for energy provision and this metabolic change is important for tumour growth. Here, we have found a link between the tumour suppressor p53, the transcription factor NF-kappaB and glycolysis. In p53-deficient primary cultured cells, kinase activities of IKKalpha and IKKbeta and subsequent NF-kappaB activity ... More
Viable cell detection by the combined use of fluorescent glucose and fluorescent glycine.
AuthorsMatsuoka H, Oishi K, Watanabe M, Kozone I, Saito M, Igimi S
JournalBiosci Biotechnol Biochem
PubMed ID14646209
'The combined use of a fluorescent glucose (2NBDG) and a fluorescent glycine (NBD-Gly) was tried for the detection of viable cells of significant foodborne pathogenic strains in addition to several Escherichia coli strains and coliforms. Thirty-five out of 41 strains showed marked uptake of 2NBDG but 6 strains were not ... More
Uptake of a fluorescent deoxyglucose analog (2-NBDG) in tumor cells.
AuthorsO'Neil RG, Wu L, Mullani N
JournalMol Imaging Biol
PubMed ID16284704
'PURPOSE: A new fluorescent analog of D -glucose was recently developed by [Yoshioka K, Takahashi H, Homma T, Sato M, Ki Bong O, Nemoto Y, Matsuoka H (1996) A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli. Biochim Biophys Acta 1289:5-9] and ... More
The histone deacetylase Sirt6 regulates glucose homeostasis via Hif1alpha.
AuthorsZhong L, D'Urso A, Toiber D, Sebastian C, Henry RE, Vadysirisack DD, Guimaraes A, Marinelli B, Wikstrom JD, Nir T, Clish CB, Vaitheesvaran B, Iliopoulos O, Kurland I, Dor Y, Weissleder R, Shirihai OS, Ellisen LW, Espinosa JM, Mostoslavsky R,
JournalCell
PubMed ID20141841
'SIRT6 is a member of a highly conserved family of NAD(+)-dependent deacetylases with various roles in metabolism, stress resistance, and life span. SIRT6-deficient mice develop normally but succumb to a lethal hypoglycemia early in life; however, the mechanism underlying this hypoglycemia remained unclear. Here, we demonstrate that SIRT6 functions as ... More
Adjuvant-induced survival signaling in clonally expanded T cells is associated with transient increases in pAkt levels and sustained uptake of glucose.
AuthorsSengupta S, Chilton PM, Mitchell TC
JournalImmunobiology
PubMed ID16325488
'Immunological adjuvants help increase the number of T cells responding to an immunizing antigen. Part of the increase is due to promotion of survival of clonally expanded T cells in the face of waning antigen load and subsequent growth-factor withdrawal. The phosphatidylinositide-3 kinase (PI3-kinase)/Akt pathway is activated upon T cell ... More
A real-time method of imaging glucose uptake in single, living mammalian cells.
AuthorsYamada K, Saito M, Matsuoka H, Inagaki N
JournalNat Protoc
PubMed ID17406637
This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent D-glucose derivative, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca2+ ... More
Evaluation of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose, a new fluorescent derivative of glucose, for viability assessment of yeast Candida albicans.
AuthorsYoshioka K, Oh KB, Saito M, Nemoto Y, Matsuoka H
JournalAppl Microbiol Biotechnol
PubMed ID8987729
A new fluorescent derivative of D-glucose, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), which had been previously developed for the analysis of glucose uptake activity by living cells, was investigated to evaluate its applicability for assaying the viability of yeast Candida albicans. Lineweaver-Burk plots showed to uptake of 2-NBDG to be competitively inhibited by D-glucose ... More
AuthorsRouach N, Koulakoff A, Abudara V, Willecke K, Giaume C,
JournalScience
PubMed ID19056987
Astrocytes provide metabolic substrates to neurons in an activity-dependent manner. However, the molecular mechanisms involved in this function, as well as its role in synaptic transmission, remain unclear. Here, we show that the gap-junction subunit proteins connexin 43 and 30 allow intercellular trafficking of glucose and its metabolites through astroglial ... More
A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli.
AuthorsYoshioka K, Takahashi H, Homma T, Saito M, Oh KB, Nemoto Y, Matsuoka H
JournalBiochim Biophys Acta
PubMed ID8605231
A novel fluorescent derivative of glucose was synthesized by reacting D-glucosamine and NBD-Cl. The TLC analysis of the reaction mixture showed the generation of a single spot with intense fluorescence (lambda Ex = 475 nm, lambda Em = 550 nm). The obtained novel fluorescent product, which was identified as 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose ... More
Molecular imaging of glucose uptake in oral neoplasia following topical application of fluorescently labeled deoxy-glucose.
AuthorsNitin N, Carlson AL, Muldoon T, El-Naggar AK, Gillenwater A, Richards-Kortum R,
JournalInt J Cancer
PubMed ID19173294
The clinical value of assessing tumor glucose metabolism via F-18 fluorodeoxyglucose (FDG) PET imaging in oncology is well established; however, the poor spatial resolution of PET is a significant limitation especially for early stage lesions. An alternative technology is optical molecular imaging, which allows for subcellular spatial resolution and can ... More
Subcellular characterization of glucose uptake in coronary endothelial cells.
AuthorsGaudreault N, Scriven DR, Laher I, Moore ED,
JournalMicrovasc Res
PubMed ID17531273
Despite all the evidence linking glucose toxicity to an increased risk of cardiovascular diseases, very little is known about the regulation of glucose uptake in endothelial cells. We have previously reported an asymmetric distribution of the GLUTs (1-5) and SGLT-1 in en face preparations of rat coronary artery endothelia [Gaudreault ... More
VX-509 (decernotinib) is a potent and selective janus kinase 3 inhibitor that attenuates inflammation in animal models of autoimmune disease.
Cytokines, growth factors, and other chemical messengers rely on a class of intracellular nonreceptor tyrosine kinases known as Janus kinases (JAKs) to rapidly transduce intracellular signals. A number of these cytokines are critical for lymphocyte development and mediating immune responses. JAK3 is of particular interest due to its importance in ... More
PINK1-associated Parkinson's disease is caused by neuronal vulnerability to calcium-induced cell death.
AuthorsGandhi S, Wood-Kaczmar A, Yao Z, Plun-Favreau H, Deas E, Klupsch K, Downward J, Latchman DS, Tabrizi SJ, Wood NW, Duchen MR, Abramov AY,
JournalMol Cell
PubMed ID19285945
Mutations in PINK1 cause autosomal recessive Parkinson's disease. PINK1 is a mitochondrial kinase of unknown function. We investigated calcium homeostasis and mitochondrial function in PINK1-deficient mammalian neurons. We demonstrate physiologically that PINK1 regulates calcium efflux from the mitochondria via the mitochondrial Na(+)/Ca(2+) exchanger. PINK1 deficiency causes mitochondrial accumulation of calcium, ... More
2-NBDG as a fluorescent indicator for direct glucose uptake measurement.
AuthorsZou C, Wang Y, Shen Z
JournalJ Biochem Biophys Methods
PubMed ID16182371
Evaluation of glucose uptake ability in cells plays a fundamental role in diabetes mellitus research. In this study, we describe a sensitive and non-radioactive assay for direct and rapid measuring glucose uptake in single, living cells. The assay is based on direct incubation of mammalian cells with a fluorescent d-glucose ... More
Glucose uptake rates of single E. coli cells grown in glucose-limited chemostat cultures.
AuthorsNatarajan A, Srienc F
JournalJ Microbiol Methods
PubMed ID11000435
To evaluate the extent to which single-cell glucose uptake rates determine the overall specific growth rate of a culture, dilute chemostat cultures of Escherichia coli BL21 were grown in defined medium under glucose limitation. The glucose uptake dynamics of the cell population was examined at the single-cell level using the ... More
Astrocytes generate Na+-mediated metabolic waves.
AuthorsBernardinelli Y, Magistretti PJ, Chatton JY
JournalProc Natl Acad Sci U S A
PubMed ID15466714
Glutamate-evoked Na+ increase in astrocytes has been identified as a signal coupling synaptic activity to glucose consumption. Astrocytes participate in multicellular signaling by transmitting intercellular Ca2+ waves. Here we show that intercellular Na+ waves are also evoked by activation of single cultured cortical mouse astrocytes in parallel with Ca2+ waves; ... More
Forward chemical genetic approach identifies new role for GAPDH in insulin signaling.
AuthorsMin J, Kyung Kim Y, Cipriani PG, Kang M, Khersonsky SM, Walsh DP, Lee JY, Niessen S, Yates JR, Gunsalus K, Piano F, Chang YT
JournalNat Chem Biol
PubMed ID17115034
Insulin and insulin-like growth factor have an essential role in growth, development and the maintenance of metabolic homeostasis, including glucose uptake from the bloodstream. Researchers have identified mutations in insulin receptors that cause severe insulin resistance, and a temperature-sensitive daf-2 (a gene encoding an insulin receptor-like protein) mutant in Caenorhabditis ... More
Fluorometric determination of glucose utilization in neurons in vitro and in vivo.
AuthorsItoh Y, Abe T, Takaoka R, Tanahashi N
JournalJ Cereb Blood Flow Metab
PubMed ID15356420
Glucose is the major energy source the adult brain utilizes under physiologic conditions. Recent findings, however, have suggested that neurons obtain most of their energy from the oxidation of extracellular lactate derived from astroglial metabolism of glucose transported into the brain from the blood. In the present studies we have ... More
Fluorescent fructose derivatives for imaging breast cancer cells.
AuthorsLevi J, Cheng Z, Gheysens O, Patel M, Chan CT, Wang Y, Namavari M, Gambhir SS
JournalBioconjug Chem
PubMed ID17444608
Breast cancer cells are known to overexpress Glut5, a sugar transporter responsible for the transfer of fructose across the cell membrane. Since Glut5 transporter is not significantly expressed in normal breast cells, fructose uptake can potentially be used to differentiate between normal and cancerous cells. Fructose was labeled with two ... More
GLUT1 deficiency links nutrient availability and apoptosis during embryonic development.
AuthorsJensen PJ, Gitlin JD, Carayannopoulos MO
JournalJ Biol Chem
PubMed ID16543226
GLUT1 is essential for human brain development and function, as evidenced by the severe epileptic encephalopathy observed in children with GLUT1 deficiency syndrome resulting from inherited loss-of-function mutations in the gene encoding this facilitative glucose transporter. To further elucidate the pathophysiology of this disorder, the zebrafish orthologue of human GLUT1 ... More
Localized glucose and water influx facilitates Cryptosporidium parvum cellular invasion by means of modulation of host-cell membrane protrusion.
Dynamic membrane protrusions such as lamellipodia and filopodia are driven by actin polymerization and often hijacked by intracellular microbes to enter host cells. The overall rate of membrane protrusion depends on the actin polymerization rate and the increase of localized cell volume. Although the signaling pathways involving actin polymerization are ... More
Thiazolidinedione-induced activation of the transcription factor peroxisome proliferator-activated receptor gamma in cells adjacent to the murine skeletal muscle: implications for fibroblast functions.
AuthorsLöhrke B, Shahi SK, Krüger B, Schmidt P, Renne U, Dietl G
JournalPflugers Arch
PubMed ID10650980
Nuclear peroxisome proliferator-activated receptor gamma (PPARgamma) is the target of antidiabetogenic thiazolidinediones (TZD). However, recent studies failed to show that TZD has an effect in vitro on insulin-regulated glucose uptake in skeletal muscles, the major site of glucose disposal. The potential effects of TZD on cells adjacent to skeletal muscles ... More
Existence of two parallel mechanisms for glucose uptake in heterotrophic plant cells.
AuthorsEtxeberria E, González P, Tomlinson P, Pozueta-Romero J
JournalJ Exp Bot
PubMed ID15911561
The implied existence of two mechanisms for glucose uptake into heterotrophic plant cells was investigated using the fluorescent glucose derivative 2-NBDG (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose), two membrane impermeable fluorescent markers (3000 mol. wt. dextran-Texas Red (d-TR) and Alexa-488), hexose carrier and endocytic inhibitors (phloridzin and wortmannin-A, respectively), and fluorescent and confocal microscopy. Both ... More
Flow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS).
AuthorsBerney M, Weilenmann HU, Egli T
JournalMicrobiology
PubMed ID16735735
The effectiveness of solar disinfection (SODIS), a low-cost household water treatment method for developing countries, was investigated with flow cytometry and viability stains for the enteric bacterium Escherichia coli. A better understanding of the process of injury or death of E. coli during SODIS could be gained by investigating six ... More
Epstein-Barr-Virus-Induced One-Carbon Metabolism Drives B Cell Transformation.
Authors
JournalCell Metab
PubMed ID31257153
GAPDH Overexpression in the T Cell Lineage Promotes Angioimmunoblastic T Cell Lymphoma through an NF-κB-Dependent Mechanism.
Authors
JournalCancer Cell
PubMed ID31447347
Reactivation of Dihydroorotate Dehydrogenase-Driven Pyrimidine Biosynthesis Restores Tumor Growth of Respiration-Deficient Cancer Cells.
Authors
JournalCell Metab
PubMed ID30449682
Store-Operated Ca2+ Entry Controls Clonal Expansion of T Cells through Metabolic Reprogramming.
Authors
JournalImmunity
PubMed ID29030115
p16(Ink4a)-induced senescence of pancreatic beta cells enhances insulin secretion.
Authors
JournalNat Med
PubMed ID26950362
High Glucose Triggers Nucleotide Imbalance through O-GlcNAcylation of Key Enzymes and Induces KRAS Mutation in Pancreatic Cells.
Authors
JournalCell Metab
PubMed ID30853214
Intestinal Epithelial and Intraepithelial T Cell Crosstalk Mediates a Dynamic Response to Infection.
Authors
JournalCell
PubMed ID28942917
Sugar-Induced Obesity and Insulin Resistance Are Uncoupled from Shortened Survival in Drosophila.
Authors
JournalCell Metab
PubMed ID32197072
Sirt2 Inhibition Enhances Metabolic Fitness and Effector Functions of Tumor-Reactive T Cells.
Authors
JournalCell Metab
PubMed ID32768387
Glutamine sensitivity analysis identifies the xCT antiporter as a common triple-negative breast tumor therapeutic target.