Trace eyeblink conditioning is associated with changes in synaptophysin immunoreactivity in the cerebellar interpositus nucleus in guinea pigs

Synaptic plasticity plays a role during trace eyeblink conditioning (TEBC). Synaptophysin (Syn) is a major integral transmembrane protein, located particularly in the synaptic vesicles, and is considered a molecular marker of synapses. In addition, Syn immunoreactivity is an important indicator of synaptic plasticity. In the present study, we used immunohistochemical techniques to assess changes in Syn expression in the cerebellar interpositus nucleus (IN) of guinea pigs exposed to TEBC and pseudoconditioning. Additionally, we analyzed the relationship between Syn immunoreactivity and the percentage of trace-conditioned responses. Guinea pigs underwent trace conditioning or pseudoconditioning. Following two, six, or ten sessions, they were perfused and the cerebellum was removed for Syn immunohistochemical evaluation. After sessions 6 and 10, a significant increase in conditioned response (CR) percentage was observed in the trace-conditioned group, with the CR percentage reaching the learning criteria following session 10. Besides, for trace-conditioned animals, the Syn expression in IN was found significantly up-regulated after session 10 compared with pseudoconditioned ones. Our data suggest that the increase in Syn expression links to synaptic plasticity changes in the cerebellar IN and provides a histological substrate in the IN relating to TEBC training. The changing trend of Syn immunoreactivity in the IN is associated with CR percentage.

Dynamic modulation of activity in cerebellar nuclei neurons during pavlovian eyeblink conditioning in mice

While research on the cerebellar cortex is crystallizing our understanding of its function in learning behavior, many questions surrounding its downstream targets remain. Here, we evaluate the dynamics of cerebellar interpositus nucleus (IpN) neurons over the course of Pavlovian eyeblink conditioning. A diverse range of learning-induced neuronal responses was observed, including increases and decreases in activity during the generation of conditioned blinks. Trial-by-trial correlational analysis and optogenetic manipulation demonstrate that facilitation in the IpN drives the eyelid movements. Adaptive facilitatory responses are often preceded by acquired transient inhibition of IpN activity that, based on latency and effect, appear to be driven by complex spikes in cerebellar cortical Purkinje cells. Likewise, during reflexive blinks to periocular stimulation, IpN cells show excitation-suppression patterns that suggest a contribution of climbing fibers and their collaterals. These findings highlight the integrative properties of subcortical neurons at the cerebellar output stage mediating conditioned behavior.