Galanin is a brain-gut peptide present in the central nervous system of vertebrates and invertebrates. The distribution of galanin-like immunoreactive perikarya and fibers in the brain of the river lamprey Lampetra fluviatilis (Agnatha) has been studied immunocytochemically by using antisera against rat and porcine galanin. Galanin-like immunoreactive perikarya were seen in the telencephalon and mediobasal diencephalon. In the telencephalon, they were present in the nucleus olfactorius anterior, nucleus basalis, and especially, in the nucleus commissurae anterioris. The diencephalon contained most of the immunoreactive neurons. They were located in the nucleus commissurae praeinfundibularis, nucleus ventralis hypothalami, nucleus commissurae postinfundibularis, nucleus ventralis thalami, and nucleus dorsalis thalami pars medius. Most of the galanin-like immunoreactive infundibular neurons showed apical processes contacting the cerebrospinal fluid. Immunoreactive fibers and terminals were widely distributed throughout the neuraxis. In the telencephalon, the richest galaninergic innervation was found in the nucleus olfactorius anterior, lobus subhippocampalis, corpus striatum, and around the nucleus septi and the nucleus praeopticus. In the diencephalon, the highest density of galanin-like immunoreactive fibers was seen in the nucleus commissurae postopticae, nucleus commissurae praeinfundibularis, nucleus ventralis hypothalami, nucleus dorsalis hypothalami, and neurohypophysis. In the mesencephalon and rhombencephalon, the distribution of immunoreactive fibers was heterogeneous, being most pronounced in a region between the nucleus nervi oculomotorii and the nucleus interpeduncularis mesencephali, in the nucleus isthmi, and in the raphe region. A subependymal plexus of immunoreactive fibers was found throughout the ventricular system. The distribution of immunoreactive neurons and fibers was similar to that of teleosts but different to those of other vertebrate groups. The possible hypophysiotropic and neuroregulatory roles of galanin are discussed.