We examined the organization of catecholaminergic neurons in the hypothalamus of the painted ray, Raja undulata, and the small-spotted dogfish, Scyliorhinus canicula, with the use of formaldehyde-induced fluorescence (FIF) methods and tyrosine hydroxylase (TH) immunohistochemistry. In both species we identified distinct populations of catecholamine-containing neurons differing in a) their immunoreactivity to antibodies against the enzyme tyrosine hydroxylase (TH), b) their fluorescence in response to FIF methods for the detection of catecholamines, and c) their relationship with the third ventricle. One population is made up of FIF-positive and TH-negative neurons (most of which are CSF [cerebrospinal fluid]-contacting) and located in two circumventricular organs, the preoptic recess organ and the organon vasculosum hypothalami. Another population comprises TH-immunoreactive (TH-IR), FIF negative neurons that are located in the suprachiasmatic nucleus and the posterior tuberculum and are not related to the third ventricle recesses. A third population of TH-IR, CSF-contacting neurons is also present in the organon vasculosum hypothalami. The existence of three catecholaminergic populations suggests differences in the metabolism of catecholamines and/or different functions. The circumventricular neurons are not associated with the hypophysis and appear to accumulate catecholamine (dopamine) obtained from exogenous sources. In both Raja and Scyliorhinus the neurointermediate lobe is innervated by TH-IR fibres originating from dopamine-synthesizing neurons of the second catecholaminergic population.