Imaging of a glucose analog, calcium and NADH in neurons and astrocytes: Dynamic responses to depolarization and sensitivity to pioglitazone

Abstract

Neuronal Ca 2+ dyshomeostasis associated with cognitive impairment and mediated by changes in several Ca 2+ sources has been seen in animal models of both aging and diabetes. In the periphery, dysregulation of intracellular Ca 2+ signals may contribute to the development of insulin resistance. In the brain, while it is well-established that type 2 diabetes mellitus is a risk factor for the development of dementia in the elderly, it is not clear whether Ca 2+ dysregulation might also affect insulin sensitivity and glucose utilization. Here we present a combination of imaging techniques testing the disappearance of the fluorescent glucose analog 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) as an indication of glycolytic activity in neurons and astrocytes. Our work shows that glucose utilization at rest is greater in neurons compared to astrocytes, and ceases upon activation in neurons with little change in astrocytes. Pretreatment of hippocampal cultures with pioglitazone, a drug used in the treatment of type 2 diabetes, significantly reduced glycolytic activity in neurons and enhanced it in astrocytes. This series of experiments, including Fura-2 and NADH imaging, provides results that are consistent with the idea that Ca 2+ levels may rapidly alter glycolytic activity, and that downstream events beyond Ca 2+ dysregulation with aging, may alter cellular metabolism in the brain.