The retrograde transport of horseradish peroxidase (HRP) and immunocytochemistry for glutamic acid decarboxylase (GAD) have been employed to examine whether local circuit neurons (LCNs) exist in the dorsal column nuclei (DCN) and whether these neurons may be GABA-ergic. Observations focused on the dorsal part of the middle cuneate nucleus (MCd), since this region has been previously shown to contain projecting neurons whose axons terminate almost exclusively in the contralateral thalamus. After large injections of HRP in the nucleus ventralis posterolateralis and surrounding structures of the feline thalamus, the majority of neurons in MCd are labeled. These represent about 89% of the neurons in MCd as counted in 40-microns frozen sections, and about 69% as counted in plastic-embedded, 2.5-microns-thick section. Unlabeled by the same injections are some medium to large neurons at the dorsal rim of MCd, and many characteristically small (mean = +/- 250 microns2) neurons at the periphery of the cell clusters formed by thalamic-projecting neurons. These small neurons represent 10-12% of the neuronal population of MCd, as counted in 40-microns-thick frozen sections, and about 30%, as counted in plastic-embedded, 2.5-microns-thick sections. Neurons in this size range are also unlabeled after injection of retrograde tracer in the pretectal area, inferior and superior colliculi, inferior olivary complex, and/or spinal cord. These injections, however, result in the labeling of neurons along the dorsal rim of MCd and/or in other regions of the cuneate nucleus. In adult, colchicine-treated cats, the use of anti-GAD serum reveals a population of labeled neurons uniformly distributed throughout the DCN. In MCd, these are small (mean = +/- 235 microns2) neurons mainly intercalated between cell clusters, and represent about 25% of the neuronal population of this nuclear subdivision as counted in plastic-embedded, 2.5-microns-thick sections. Labeled processes densely infiltrate the cell clusters, and labeled varicosities appear to cover the soma and dendrites of unlabeled neurons. At the electron-microscopic level, most labeled profiles contain vesicles and correspond to F boutons usually involved in "axoaxonic" contacts with terminals of dorsal root afferent and presynaptic to dendrites. Other vesicle-containing, GAD-positive endings seem to correspond to the P boutons described by Ellis and Rustioni (1981) and are believed to be, at least in part, of dendritic origin. It is suggested that GAD-positive neurons are GABA-ergic LCNs and that these can mediate both pre- and postsynaptic inhibition.(ABSTRACT TRUNCATED AT 400 WORDS)