PhosphoRiboTrap sequencing data of chemogenetic MCH neuron activation

We report phosphoRiboTrap experiments result which aim to explore the nature of the cell types in the Median Eminence (ME) regulated as a consequene of chemogenetic MCH neuron activation. Control and MCH-hM3Dq mice were 12 hrs-fasted and i.p. injected with CNO (3 mg/kg), Arcuate (ARC) and median eminence expat were extracted for preciptation of S6-marked ribosomes from both groups of mice. Extracted RNA from Immunoprecipitated ribosomes (IP) and total tissue (ARC+ME) from each mouse were subjected to deep mRNA sequencing. By analyzing the overlap of genes enriched in the IP/Input of MCH neuron activated mice with previously identified cell types using single cell mRNA sequencing of cells in the mediobasal hypothalamus (Campbell et al., 2017), we identified gene clusters which were activated upon MCH neuron activation. Overall design: 7 control and 5 MCH-hM3Dq mice were subjected to 12 hrs and 1 hr CNO injection. Mice were acutely decapitated. ARC and ME from hypothalamus tissue were cut with brain matrix and collected on dry ice and frozen in liquid nitrogen and stored in -80 freezers. All the samples were subjected to PhosphoRiboTRAP protocol (see details in protocol part) and both IP and Input were harvested and concentration were measured by Qbid machine. Those samples were deep mRNA sequenced for further analysis.