Substance P-like immunoreactivity was examined in the Limulus lateral eye and photocerebrum by the unlabeled antibody peroxidase-antiperoxidase method. Small-diameter immunoreactive fibers innervate the corneal epidermis of the lateral eye and appear to be part of a generalized epidermal innervation. No immunoreactive neurons or fibers were found in the ventral, median, or lateral optic nerves nor in the lamina or chiasma. The optic medulla contains neurons with immunoreactive perinuclear caps in the ganglion cell layer and fibers of undetermined origin in the posterolateral regions of the neuropil. The central body contains many immunoreactive fibers in its neuropil, some or all of which arise from neurons adjacent to its anteromedial tips. Immunoreactive neurons within the curve of the central body give rise to processes which join the central neuropil of the photocerebrum. The immunoreactive innervation of the corpora pedunculata includes processes which terminate amont Kenyon cell somata, and processes in the peduncular neuropil, all of which arise from a bilateral cluster of neurons along the midline. Attempts to investigate the source of the immunoreactive fibers using cobalt impregnation of the severed circumesophageal connective have revealed two additional innervations of the corpora pedunculata, one from raphe neurons along the midline and one from lateral portions of the midline ganglion adjacent to but separate from that portion containing the immunoreactive neurons. The results demonstrate that immunocytochemical methods are another powerful tool for the study of neuronal pathways in invertebrates.