Molecular profiling of hypothalamic leucine-sensing neurons via PhosphoTRAP
To identify the molecular signature of leucine-activated neurons in the MBH, we adopted the PhosphoTRAP assay following parenchymal injection of leucine into the MBH.  Neurons activated following MBH leucine injection (and expressing the neuronal activation marker cFos) co-express Ser240/244-phosphorylated ribosomal S6 protein , allowing selective immunoprecipitation-based capture (TRAP) and RNA sequencing of polysomes from leucine-activated neurons. Differential expression analysis between IP and input samples generated a list of enriched transcripts in leucine-activated neurons