The extent to which the cells of origin of the cerebellar nucleocortical pathway are immunopositive for gamma-aminobutyric acid (GABA) was investigated in four cats using retrograde labelling of nucleocortical neurons in combination with immunocytochemistry. Neurons were retrogradely labelled by injection of fluorescent (coumarin)-tagged latex microspheres into the c2 zone in the rostral part of the paramedian lobule. The zone was identified electrophysiologically by the characteristics of the climbing fibre responses evoked on the cerebellar surface by percutaneous stimulation applied to the left and right forepaws in pentobarbitone-anaesthetized animals. Sections of the cerebellum containing the retrogradely labelled neurons were processed for GABA immunocytochemistry using a fluorescent (rhodamine)-tagged immunoglobulin. When viewed with epifluorescence microscopy and appropriate filter blocks the retrogradely labelled nucleocortical neurons could be visualized in the same sections as the GABA-immunopositive neurons. Almost all of a total of 254 labelled nucleocortical neurons were located in nucleus interpositus posterior, where a total of 711 GABAergic neurons were also found. None of these cells contained coumarin-tagged beads and displayed immunoreactivity for GABA (i.e. none was double-labelled). When compared by area of their cell body, the nucleocortical and GABA-immunopositive neurons appeared to form two partially overlapping populations. The mean cell area of the nucleocortical neurons was 620 +/- 233 microns2 (SD), whereas the GABA-immunopositive neurons were much smaller, with a mean cell area of 220 +/- 115 microns2. The results suggest that GABA does not play a major role in the nucleocortical pathway to the c2 zone of the rostral paramedian lobule of the cat cerebellum.