Projection from the cerebellar lateral nucleus to precerebellar nuclei in the mossy fiber pathway is glutamatergic: a study combining anterograde tracing with immunogold labeling in the rat.

The pontine nuclei (PN) and the nucleus reticularis tegmenti pontis (NRTP) are sources of an excitatory projection to the cerebellar cortex via mossy fibers and a direct excitatory projection to the cerebellar nuclei. These precerebellar nuclei, in turn, receive a feedback projection from the cerebellar nuclei, which mostly originate in the lateral nucleus (LN). It has been suggested that the feedback projection from the LN partially uses gamma-aminobutyric acid (GABA) as a transmitter. We tested this hypothesis by using a combination of anterograde tracing (biotinylated dextran amine injection into the LN) and postembedding GABA and glutamate immunogold histochemistry. The pattern of labeling in the PN and the NRTP was compared with that of cerebellonuclear terminals in two other target structures, the parvocellular part of the nucleus ruber (RNp) and the ventromedial and ventrolateral thalamus (VM/VL). The projection to the inferior olive (IO), which is known to be predominantly GABAergic, served as a control. A quantitative analysis of the synaptic terminals labeled by the tracer within the PN, the NRTP, and the VL/VM revealed no GABA immunoreactivity. Only one clearly labeled terminal was found in the RNp. In contrast, 72% of the terminals in the IO were clearly GABA immunoreactive, confirming the reliability of our staining protocol. Correspondingly, glutamate immunohistochemistry labeled the majority of the cerebellonuclear terminals in the PN (88%), the NRTP (90%), the RNp (93%), and the VM/VL (63%) but labeled only 5% in the IO. These data do not support a role for GABAergic inhibition either in the feedback systems from the LN to the PN and the NRTP or within the projections to the RNp and the VM/VL.