An immunohistochemical method that utilizes carbodiimide as a fixative and antisera directed against histamine was applied to investigate the location of histamine in the rat pineal complex. Numerous histamine-immunoreactive cell bodies were observed in different subdivisions of the tuberomammillary nucleus of the posterior hypothalamus, and a few cell bodies were present in the posterior and dorsal part of the periventricular hypothalamic nucleus. Histamine-immunoreactive fibers were observed to leave the posterior hypothalamus in various directions of which one dorsally projecting tract was followed in the periventricular area of the caudal diencephalon to the epithalamus. Several histamine-immunoreactive nerve fibers of this tract continued through the posterior commissure directly into the deep pineal gland. A few immunoreactive fibers were also observed in the habenular commissure. In midsagittal sections, histamine-immunoreactive nerve fibers were observed to enter the pineal stalk from the deep pineal gland. Most of histamine-immunoreactive fibers in the stalk continued towards the superficial pineal gland, but their number decreased in more distal locations of the stalk, indicating that some fibers terminate in the stalk as well. A few fibers were found to terminate in the most rostral part of the superficial pineal gland. The immunoreactive nerve fibers in the epithalamus and pineal complex were endowed with prominent varicosities. Taken together, these results indicate that histaminergic nerve fibers, originating from the posterior hypothalamus, project to the pineal complex of the rat. Histamine must therefore be considered a putative neurotransmitter contained in the central innervation of the pineal gland, but its function in pineal physiology has so far not been elucidated.