Electron microscopic study of the rubrocerebellar projection in the cat

Rubral neurons sending axons to the cerebellar anterior interpositus nucleus (AIN) in the cat were identified light microscopically by labeling them with horseradish peroxidase (HRP). The synaptic organization of these rubral neurons and of their afferents from the cerebral motor cortex and the AIN was also analyzed electron microscopically by combined anterograde degeneration and retrograde HRP-labeling techniques. In the light microscopic study, either HRP or a mixture of HRP and kainic acid was injected into the AIN. Both of the injections resulted in retrograde labeling of rubrocerebellar projection neurons in the red nucleus on the contralateral side. The labeled neurons were distributed throughout the rostrocaudal extent of the red nucleus: some lay in clusters. Most labeled neurons were small to medium-sized, although some were large. The injection of HRP into the AIN also resulted in anterograde labeling of cerebellorubral projection fibers terminating in a wider area of the red nucleus on the contralateral side of the injection, whereas the injection of a mixture of HRP and kainic acid showed no anterograde labeling of fibers or terminals. In one set of electron microscopic observations, HRP injections into the AIN were combined with ablation of the motor cortex. Degenerating axon terminals were occasionally found to synapse with both dendrites and neuronal somata labeled with HRP retrogradely. In another set of electron microscopic observations, a mixture of HRP and kainic acid was injected into the AIN in order to label rubrocerebellar projection neurons retrogradely and to bring about degeneration in the cerebellorubral projection fibers anterogradely. Abundant degenerating axon terminals were observed to make axosomatic synaptic contacts with rubral neurons labeled with HRP retrogradely and also with unlabeled rubral neurons. These results indicate that cerebrorubrocerebellar and rubrocerebellorubral monosynaptic circuitries exist which constitute one of the cerebrocerebellar linkages, as well as those linkages via the inferior olivary complex and the pontine nuclei.