Excitatory amino acids as neurotransmitters of cortical and cerebellar projections to the red nucleus: an immunocytochemical study in the guinea pig.

We combined a retrograde labeling technique with peroxidase immunocytochemistry to verify whether cortical and cerebellar neurons projecting to the red nucleus (RN) contain high concentrations of glutamate and aspartate as possible neurotransmitters. Injections of a tracer, colloidal gold-labeled enzymatically inactive horseradish peroxidase conjugated to wheatgerm agglutinin, into the RN of adult guinea pigs produced retrograde labeling of layer V cortical neurons, with a large predominance in the ipsilateral hemisphere. Corticorubral neurons were located in the granular parietal cortex (Gr), agranular frontal cortex (Ag), agranular cingulate cortex (Cg), and retrobulbar cortex (Rb). Large numbers of retrogradely labeled neurons were concentrated in contralateral interpositus and dentate cerebellar nuclei. We found the majority of corticorubral neurons to be immunostained by antibodies raised in rabbits against glutamate or aspartate conjugated to invertebrate hemocyanin by glutaraldehyde, supporting the hypothesis that excitatory amino acids are neurotransmitters of corticorubral projections. With either antiserum, immunostaining was found in 58-72% of corticorubral neurons in Ag and Gr; higher percentages were observed in Rb (80-85%) and Cg (up to 96%). Cross-sectional area measurements indicated that the perikarya of corticorubral neurons were larger in Ag and Gr than in Rb and Cg; in each area, soma size values of immunopositive corticorubral neurons tended to be larger than those of immunonegative ones. In the cerebellar nuclei, virtually all retrogradely labeled neurons were immunostained by glutamate and aspartate antisera, suggesting that excitatory amino acids might also be considered as possible neurotransmitters for cerebellorubral projections.