Dynamic monitoring of cytosolic glucose in single astrocytes

Abstract

It is becoming increasingly clear that astrocytes are no longer playing a subservient role to neurons in the central nervous system (CNS), and that these cells are being considered as active communication integrators. They respond to neurotransmitters by the regulated release of gliotransmitters. The delay between neurotransmitter activation and the release of gliotransmitters from astrocytes is in the time-domain of subseconds, much slower than the submillisecond synaptic delay. Astrocytes also control microcirculation and provide metabolic support for neurons. However, the dynamics of their energy metabolic response to neurotransmitter application is not known. We here used a FRET glucose nanosensor to dynamically measure the cytosolic glucose concentration in single astrocytes. We show that following the adrenaline or noradrenaline stimulation the availability of cytosolic glucose is increased promptly after stimulation with a time-constant of 116.7 s and 115.9 s, respectively. A decline in cytosolic glucose concentration with a time-constant of 50.7 s was observed during glutamate and 16.7 s during lactate addition to astrocytes, when these were bathed in the presence of extracellular glucose-containing solution, likely reflecting predominant glucose engagement in glycogen synthesis. In contrast, in the glucose-free extracellular solution, glutamate application to astrocytes resulted in a slow increase in cytosolic glucose concentration, consistent with the view that glutamate may be an alternative energy source in hypoglycemic conditions. We conclude that astrocytic cytosolic glucose metabolism responds in the time-domain of tens of seconds, which is slower compared to the whole brain functional magnetic resonance imaging measurements of the local intravascular hemodynamic response.