Isotope-Based Quantitation of Uptake, Release, and Metabolism of Glutamate and Glucose in Cultured Astrocytes

Protocols are described for measurement in primary cultures of astrocytes of unidirectional fluxes of glutamate (influx and efflux), glutamate metabolism to glutamine or CO₂ , glucose influx, glycolysis, pyruvate dehydrogenation, oxidative metabolism of glucose, pyruvate carboxylation, glycogen synthesis, and glycogenolysis. References are made to the in vivo situation, and the importance of using metabolically competent cultures is emphasized.