Under direct visualization, minute amounts of horseradish peroxidase were injected by controlled air pressure through glass micropipettes into the inferior olive of the albino rat. Well localized unilateral injections were obtained, without damage to other brainstem areas or spread into the adjacent reticular formation. After 24 to 36 hours, the entire brain and spinal cord were examined by light microscopy for labelled neurons supplying afferents to the inferior olive.With few exceptions, labelled somata rostra1 to the inferior olive were found ipsilateral to the injection site, the greatest number occurring in the subparafascicular nucleus and central gray substance around the caudal third ventricle, in the N. of Forel's field, Nn. of Darkschewitsch and Cajal, and adjacent reticular formation, and in the Edinger-Westphal nucleus. Labelled cells were also noted in the sensorimotor cortex, red nucleus, peri-aqueductal gray and all subdivisions of the pretectal complex. Caudal to the olive, labelled neurons were located primarily on the contralateral side in the lateral reticular nucleus, dorsal column nuclei and nucleus proprius of the spinal cord. At the level of the olive, numerous labelled cells occurred bilaterally, but mainly contralateral to the injection site in the lateral and interpositus nuclei of the cerebellum. A smaller number were seen in the N. prepositus hypoglossi and in the medial and spinal subdivisions of the vestibular complex ipsilaterally, in the N. spinal tract of V contralaterally, and in the gigantocellular reticular nucleus bilaterally. A few neurons in the N. raphe obscurus were also labelled. No positive cells were found in the caudate nucleus, globus pallidus, locus coeruleus, or medial cerebellar nucleus.The results are compared and contrasted, region by region, with previous studies of afferent olivary pathways and, where possible, correlations are made with physiologic data. studies over the past ten years, and during this time many theories have emerged. These have been amply reviewed by Armstrong ('74) and Chan-Palay ('77) and will not be further recounted here. What has been lacking, however, is a more precise determination of the various sources of afferents to the inferior olive by which this very important pathway might be influenced.Early anterograde degeneration studies with Marchi and reduced silver techniques began the piecemeal assembly of olivary afferents, but even discrete lesions ran the risk of destroying fibers of passage as well as cell bodies, and there was often controversy over exactly what constituted terminal degeneration. Autoradiography has overcome most of these difficulties, but the problem of limitation of injection sites remains, and this method still enables determination of only one afferent source a t a time, although it is extremely useful in defining precisely which regions of a nucleus like the olive receive afferents from a particular source. The method of retrograde axonal transport of horseradish peroxidase (HRP) as developed by Krist...