Amylin is a recently discovered 37 amino acid peptide which is co-secreted from the pancreas with insulin and acts to modulate carbohydrate metabolism. Recently, high-affinity binding sites for [125I]rat amylin have been identified in the rat central nervous system. These sites also have high affinity for the structurally related peptides calcitonin gene-related peptide and salmon calcitonin. In the present study we have used in vitro autoradiography to map the distribution of these [125I]rat amylin binding sites in rat brain. High to moderate levels of binding were present in mid-caudal accumbens nucleus, fundus striati and parts of the bed nucleus of the stria terminalis and substantia inominata. This binding extended caudally into parts of the amygdalostriatal transition zone and the central and medial amygdaloid nuclei. High to moderate levels of binding also occurred in much of the hypothalamus including the medial preoptic, dorsomedial hypothalamic and medial tuberal nuclei as well as the ventrolateral subnucleus of the ventromedial hypothalamic nucleus. Other regions of high level binding included the subfornical organ, the vascular organ of the lamina terminalis, area postrema, locus coeruleus, dorsal raphe and caudal parts of the nucleus of the solitary tract. The subfornical organ, vascular organ of the lamina terminalis and area postrema, which display some of the highest binding densities, lack a patent blood-brain barrier and thus could be responsive to blood-borne amylin. In conclusion we have mapped, in detail, the distribution of amylin binding sites in rat brain. The location of binding is consistent with potential roles for these sites in appetite, fluid and electrolyte homeostasis, autonomic function and regulation of mood.