The hypothalamic neurotransmitter dopamine (DA) regulates pituitary secretion of the glucoregulatory hormones, growth hormone (GH) and adrenocorticotropin (ACTH). The glucose antimetabolite, 2-deoxy-D-glucose (2DG), elicits expression of the proto-oncogene product Fos, which is expressed in hypothalamic structures where DA is synthesized. These studies utilized dual-label immunocytochemistry to determine whether discrete DA neuron populations in this region of the brain exhibit Fos immunoreactivity (-ir) in response to glucopenia. Ovariectomized female rats implanted s.c. with exogenous estradiol or vehicle were injected with 2DG (400 mg/kg, i.p.) or saline, and sacrificed 2 h later. Whereas Fos-ir was negligible after saline administration, 2DG induced expression of Fos-ir by TH-ir neurons in the paraventricular (PVN), periventricular (Pe) and arcuate nuclei (ARC), and in the anterior hypothalamic area (AHA). TH-ir neurons in the zona incerta did not express Fos-ir following 2DG. Although mean numbers of co-labeled neurons in the Pe, PVN and AHA did not differ between estradiol- and non-steroid-treated rats, the former group exhibited significantly higher numbers of TH-positive plus Fos-positive neurons in the ARC in response to 2DG. These results reveal the functional responsiveness of discrete DA neuron populations to glucoprivation, and indicate that estradiol enhances cellular accumulation of Fos-ir by ARC DA neurons during this metabolic challenge.