Gene expression profiling of Vglut2 neurons in the vestibular nucleus (VN)

Vestibular nuclei (VN) are critical for the processing of movement input, and motion-induced activation of VN neurons recapitulates MS-related signs. However, the genetic identity of VN neurons mediating MS-related autonomic and aversive responses remains unknown. Here, we identify diferent markers of glutamatergic vestibular neuronal populations using the RiboTag apparoach. Punches containing the vestibular nuclei (VN) of Vglut2Cre mice injected with the RiboTag viral vector (AAV-DIO-Rpl22·HA) were homogenized and incubated with anti-hemagglutin (HA) antibodies and protein A/G magnetic beads to isolate the polysome-associated mRNAS of VGLUT2 neurons in the VN. To identify differentially expressed transcripts, RNA from the immunoprecipitates was extracted, amplified, labelled and hybridized to MouseRef-8 v2 expression beadchips.