A procedure for the simultaneous visualization of two anterograde and different retrograde fluorescent tracers. Application to the study of the afferent-efferent organization of thalamic anterior intralaminar nuclei.

The present report describes a method for the simultaneous visualization, in the same structure, of two different sets of afferent pathways and the neurons of origin of some efferent projections. This method has been applied in the cat for studying, in the thalamic anterior intralaminar nuclei, the topographical relationships of afferent arising from the spinal cord and deep cerebellar nuclei with neurons projecting to different cortical areas. Spino- and cerebello-thalamic terminals were anterogradely labeled by injections of the fluorescent dyes fast blue (FB) and 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI) in the spinal cord and cerebellum. Thalamo-cortical neurons were retrogradely labeled by injections of fluorescent tracers in the precruciate and anterior suprasylvian cortices. The findings show that spinal and cerebellar afferent fibers and the cells of origin of intralaminocortical projections are organized in a clear modular manner and indicate that the method used here is suitable for analyzing simultaneously, in light microscopy, multiple input-output interrelationships of a single structure.