Gene expression profiling of Calca neurons in the parabrachial nucleus (PBN)

Calca-expressing neurons in the parabrachial nucleus (PBN) play a major role in regulating appetite and transmitting real or potential threat signals. Here, we reveal transcripts specific or highly enriched in this neuronal population by using mouse genetics and a conditional viral ribosome-tagging approach to phenotype these neurons. Punches containing the parabrachial nucleus (PBN) of Calca-Cre 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 Calca neurons in the PBN. To identify differentially expressed transcripts, RNA from the inputs and immunoprecipitates was extracted, amplified, labelled, and hybridized to MouseRef-8 v2 expression beadchips.