Abstract
The glycolytic rate is sensitive to physiological activity, hormones, stress, aging, and malignant transformation. Standard techniques to measure the glycolytic rate are based on radioactive isotopes, are not able to resolve single cells and have poor temporal resolution, limitations that hamper the study of energy metabolism in the brain and other organs. A new method is described in this article, which makes use of a recently developed FRET glucose nanosensor to measure the rate of glycolysis in single cells with high temporal resolution. Used in cultured astrocytes, the method showed for the first time that glycolysis can be activated within seconds by a combination of glutamate and K+, supporting a role for astrocytes in neurometabolic and neurovascular coupling in the brain. It was also possible to make a direct comparison of metabolism in neurons and astrocytes lying in close proximity, paving the way to a high-resolution characterization of brain energy metabolism. Single-cell glycolytic rates were also measured in fibroblasts, adipocytes, myoblasts, and tumor cells, showing higher rates for undifferentiated cells and significant metabolic heterogeneity within cell types. This method should facilitate the investigation of tissue metabolism at the single-cell level and is readily adaptable for high-throughput analysis.
Highlights
The metabolic rate defines the speed at which the body burns fuel
The metabolic rate of an individual cell can vary through time: for instance, adipocytes increase their rate of glucose uptake by up to ten-fold in response to insulin, whereas neurons may raise their energy demand by larger factors in response to electrical stimulation
Theory The concentration of glucose can be measured in single cells using FRET nanosensors (Fehr et al, 2003; Deuschle et al, 2005; Takanaga et al, 2008; Takanaga and Frommer, 2010)
Summary
The metabolic rate defines the speed at which the body burns fuel. Each cell is characterized by a specific metabolic rate, ranging from low for quiescent cells like fibroblasts to very high for active muscle cells and neurons. The metabolic rate of an individual cell can vary through time: for instance, adipocytes increase their rate of glucose uptake by up to ten-fold in response to insulin, whereas neurons may raise their energy demand by larger factors in response to electrical stimulation. Cancer cells show higher metabolic rates than their normal counterparts, a phenomenon involved in tumor progression and instrumental for the purposes of diagnosis, staging and prognosis of this disease (Vander Heiden et al, 2009; Yeluri et al, 2009)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
