Activation of septal GABA receptors impairs learning and memory and this effect likely involves an influence on the hippocampus. We found previously that the memory-impairing effects of septal infusions of the GABA agonist muscimol are reversed by hippocampal infusions of glucose and suggested that glucose reverses these deficits by increasing hippocampal acetylcholine (ACh). In this study, we report that septal infusions of muscimol produce dose-dependent decreases in ACh levels in hippocampal dialysates. Importantly, increasing glucose levels in the hippocampus elevates hippocampal extracellular ACh levels in rats given septal infusions of muscimol, but not in rats given vehicle. Thus, glucose increases hippocampal extracellular ACh levels when the ACh system is inhibited, an effect that likely contributes to the effects of glucose on memory.