The development of substance P-like immunoreactivity (SPLI) in the goldfish brain was studied by means of the indirect peroxidase-antiperoxidase technique and an antibody to substance P. By 80 h after fertilization, the first SPLI-cell bodies appear in the ventricular zone of the future diencephalon and the first SPLI-fibers appear in the olfactory bulbs. Two days after hatching (which occurs at 100 h after fertilization), SPLI fibers connecting the olfactory bulbs and hypothalamus are seen. In the optic tectum SPLI-fibers appear for the first time 5 days after hatching. In the brain stem, SPLI-cell bodies appear in juvenile animals 40 days after hatching. The highest number and intensity of SPLI-cell bodies and fibers are found in the area postrema. SPLI-cell bodies are also seen in the gustatory nucleus, nucleus ambiguus, reticular formation of the medulla, dorsal motor nucleus of the vagus and commissural nucleus of Cajal. The significant information gained from the present study is: 1. The rostro-caudal sequence in which the SPLI appears in the developing nuclei of the goldfish brain 2. The reduction of SPLI-cell bodies in some nuclei with age Thus, in the brain stem, SPLI-cell bodies that were labeled in juvenile goldfish were not seen in adults. This might be due to changes in the rate of axonal transport, changes of the SP phenotype during development or cell death. The developmental sequence and relative timing in which SPLI-cell bodies appear in the goldfish, rat and mice are similar.