Using a rabbit endothelin-1 (ET-1) antiserum together with a goat-anticholineacetylase antiserum or a mouse anti-somatostatin antiserum it was possible by means of double immunolabelling procedures to demonstrate ET-like immunoreactivity in striatal nerve cell bodies of the rat, which were shown to contain either cholineacetylase or somatostatin immunoreactivity. Absorption studies with ET-3, ET-1 or big ET-1 indicated that the ET-like immunoreactivity was ET-3 like. In agreement the radioimmunoassay showed that ET-3-like immunoreactivity was present in higher concentrations than ET-1-like immunoreactivity in the neostriatum and other brain areas. Characterization by reversed phase HPLC revealed, however, that a major portion of the neostriatal ET-3-like immunoreactivity was not identical to ET-3. The nature of neuronal ET in the rat may thus be more complicated than hitherto assumed.