Surgical ablations of telencephalon or telencephalon plus habenular nuclei (HBN) have been performed for studying anatomical connections of the telencephalo-habenulo-interpeduncular system of the goldfish. The results of the ultrastructural studies confirmed the presence of massive telencephalo-habenular and habenulo-interpeduncular projections and, in addition, demonstrated a minor direct telencephalo-interpeduncular connection. Immunohistochemical localization of substance P in the interpeduncular nucleus (IPN) after telencephalic ablation failed to demonstrate the involvement of this neurotransmitter in the direct telencephalo-interpeduncular projection, while telencephalon plus HBN ablation reduced substance P immunoreactivity in the IPN. The different distribution of degenerating terminals converging on the IPN from the HBN and from the telencephalon suggested a subnuclear organization of this area, as described in mammals.