Thirst regulation involves several brain regions and Glutamatergic/GABAergic mechanisms. Imaging those noninvasively could make a big impact on understanding brain circuitry in-vivo. A recently proposed method termed overlap-resolved CEST (orCEST) was suggested for mapping both Glutamate and GABA in the brain with increased specificity. Here, orCEST is used to investigate changes in these neurotransmitters in the rat brain in vivo upon water deprivation. Decreases in GABA and Glutamate in hypothalamic preoptic areas and corpus callosum, respectively, were observed in the Water Deprived group. This bodes well for future applications of orCEST for studying neurotransmission in-vivo.