The distribution of radiolabeled neurons in the brain stem of Lampetra fluviatilis was studied following unilateral injections of (3H)D-aspartate in the rostral spinal cord. After survival periods of 1-3 days, labeled perikarya were present within and nearby the posterior, middle, and anterior rhombencephalic reticular nuclei and in the mesencephalic reticular nucleus. The highest number of (3H)D-aspartate labeled cell bodies were present in the posterior rhombencephalic reticular nucleus. The labeled reticulospinal neurons were distributed mainly ipsilateral to the injection site and included the giant Müller cells as well as medium-sized and small neurons. Contralateral labeling occurred in cell bodies scattered along the lateral margin of the rhombencephalic reticular formation, the most rostral of these contralaterally projecting neurons being the Mauthner cell. The (3H)D-aspartate labeling correlates with previous electrophysiological studies showing that lamprey reticulospinal neurons utilize excitatory amino acid transmission.