A simplified approach to retrograde/anterograde axonal labeling using combined injections of horseradish peroxidase and ibotenic acid.

Combined injections of ibotenic acid and horseradish peroxidase (HRP) were made into the region of the mouse ventrobasal thalamus that is related to the large mystacial vibrissae. Examination 4 and 5 days later of the corresponding area of the primary somatosensory cortex (i.e., barrel cortex), in thick and in thin sections, showed it to contain numerous corticothalamic projection cells the somata, dendrites and axons of which were densely labeled by the retrograde transport of HRP. Analysis of serial thin sections showed that thalamocortical axon terminals, which had degenerated in response to the injection of ibotenic acid, formed approximately 20% of the asymmetrical synapses in barrel cortex. The fine structure and distribution in cortex of degenerating thalamocortical axon terminals and of intrinsic HRP-labeled corticothalamic axon terminals were identical to those reported in previous studies in which the injection of HRP into the thalamus was combined with the making of electrolytic lesions. This indicates that injecting ibotenic acid is an effective replacement for electrolytic lesioning of the thalamus. The combined injection of ibotenic acid and HRP represents a new and efficient approach for studying reciprocal projection pathways.