Superovulated mouse oocytes and embryos were isolated and incubated with nontracer concentrations of 2-deoxyglucose, an analog of glucose, which is transported into the cells by glucose transporter and phosphorylated by hexokinase to 2-deoxyglucose 6-phosphate; it is not rapidly metabolized further, and accumulates in the cytosol. Using our non-radiometric and enzymatic microassay method, we determined 2-deoxyglucose and 2-deoxyglucose 6-phosphate amounts in individual oocytes and preimplantation embryos after the incubation. Hexokinase activity increased continuously and exponentially during development from follicular oocytes to blastocysts. Endogenous glucose and glucose 6-phosphate decreased precipitously from follicular oocytes to unfertilized and ovulated oocytes. Fertilization induced rapid increases in glucose and glucose 6-phosphate concentrations, which increased exponentially thereafter during embryonic development. 2-Deoxyglucose incorporation and 2-deoxyglucose 6-phosphate formation were undetectable in unfertilized oocytes. However, when cumulus-oocyte complexes were incubated with 2-deoxyglucose, 2-deoxyglucose was incorporated into cumulus cells surrounding follicular oocytes and transported via the gap junctions into the follicular oocytes. Fertilization triggered facilitative 2-deoxyglucose transport in one-cell embryos, and the capacities of 2-deoxyglucose incorporation and 2-deoxyglucose 6-phosphate formation developed along with the maturation of preimplantation embryos.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1095/biolreprod50.2.266 | DOI Listing |
STAR Protoc
December 2024
Department of Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan. Electronic address:
2-deoxy-D-glucose (2DG) is a glucose analog converted to 2-deoxy-D-glucose-6-phosphate (2DG-6P) by hexokinase in glycolysis. While 2DG commonly measures glucose uptake, 2DG-6P detects glucose utilization. Here, we present a protocol to measure glucose utilization in various tissues after entering a mouse's body using radiolabeled 2DG.
View Article and Find Full Text PDFMicrobiol Spectr
August 2024
Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, Leuven, Belgium.
Unlabelled: The genes, and , encode for 2-deoxyglucose-6-phosphate phosphatases. These enzymes of the haloacid dehalogenase superfamily are known to utilize the non-natural 2-deoxyglucose-6-phosphate as their substrate. However, their physiological substrate and hence their biological role remain elusive.
View Article and Find Full Text PDFNeurochem Res
December 2024
Centre for Biomolecular Interactions Bremen, Faculty 2 (Biology/Chemistry), University of Bremen, P.O. Box 330440, 28334, Bremen, Germany.
The glucose analogue 2-deoxyglucose (2DG) has frequently been used as a tool to study cellular glucose uptake and to inhibit glycolysis. Exposure of primary cultured astrocytes to 2DG caused a time- and concentration-dependent cellular accumulation of 2-deoxyglucose-6-phosphate (2DG6P) that was accompanied by a rapid initial decline in cellular ATP content. Inhibitors of mitochondrial respiration as well as inhibitors of mitochondrial uptake of pyruvate and activated fatty acids accelerated the ATP loss, demonstrating that mitochondrial ATP regeneration contributes to the partial maintenance of the ATP content in 2DG-treated astrocytes.
View Article and Find Full Text PDFImmun Inflamm Dis
November 2023
Department of Burn and Plastic Surgery, General Hospital of Northern Theater Command, Shenyang, Liaoning, China.
Background: Insulin resistance (IR) is considered as a major factor initiating type 2 diabetes mellitus and can lead to a reduction in glucose uptake that mainly occurs in the liver. Astragalus polysaccharide (APC), extracted from the traditional Chinese medicine, has been recorded to suppress IR. However, the underlying mechanism remains inadequately explored.
View Article and Find Full Text PDFPLoS Genet
August 2022
Université Paris Cité, CNRS, Institut Jacques Monod, Paris, France.
2-deoxyglucose is a glucose analog that impacts many aspects of cellular physiology. After its uptake and its phosphorylation into 2-deoxyglucose-6-phosphate (2DG6P), it interferes with several metabolic pathways including glycolysis and protein N-glycosylation. Despite this systemic effect, resistance can arise through strategies that are only partially understood.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!