We use the method of retrograde transport of colloidal gold with silver intensification to map locations of brainstem neurons projecting to various nuclei of the medial and lateral thalamus (Menetrey, Histochemistry 83, 391-395, 1985; Seeley and Field, Brain Res. 449, 177-191, 1988). In rats injections of less than 1.0 microliters of a gold-wheatgerm agglutinin-horseradish peroxidase complex were restricted to the centrum medianum, centralis lateralis, medialis dorsalis, zona incerta, ventrobasal thalamic complex, or medial or lateral hypothalamus. Injections in the centralis lateralis, with some involvement of centrum medianum and medial medialis dorsalis, labeled numerous neurons throughout the rostrocaudal extent of the midbrain periaqueductal gray, and adjacent reticular formation, dorsal raphe nucleus and substantia nigra. These labeled neurons were distributed bilaterally, but with a preponderance ipsilaterally. Numerous neurons in the deep layers of the ipsilateral superior colliculus and in the anterior pretectal nucleus were also labeled. Many neurons in the pontomedullary raphe nuclei (including nucleus raphe magnus), locus coeruleus and dorsolateral parabrachial nuclei, and in the pontomedullary reticular formation, were labeled bilaterally. Fewer were seen bilaterally in the lateral reticular nucleus and nucleus of the solitary tract, with relatively few in the dorsal aspect of the trigeminal nucleus caudalis. Injections restricted largely to the centrum medianum labeled fewer cells in the brainstem, but with similar distributions. An injection restricted to the centralis lateralis also resulted in a similar distribution of labeled neurons in midbrain and nucleus raphe magnus but did not label neurons at more caudal medullary levels. An injection restricted to zona incerta labeled numerous neurons in a distribution similar to that following medial thalamic injections, except that many more were observed in the contralateral sensory trigeminal nucleus and in the dorsal column nuclei. Injections of the ventrobasal thalamic nucleus labeled many neurons in the trigeminal and dorsal column nuclei, but many fewer neurons in the midbrain, periaqueductal gray and reticular formation compared with medial thalamic injections. Labeled neurons were also seen in the superior and inferior colliculi, due presumably to involvement by the injection of the lateral posterior and magnocellular medial geniculate nuclei, respectively. Injections in the lateral hypothalamus labeled numerous neurons in a distribution similar to that seen following medial thalamic injections.(ABSTRACT TRUNCATED AT 400 WORDS)