Sciatic nerve injury-responsive differential mRNA profile and ribosome association efficiency of injury-induced mRNA

After nerve injury in the pheripheral nervous system, axon regeneration occurs through transcriptomic changes in neuronal cell bodies. To investigate which protein-coding transcripts are up-regulated and directed to ribosomes or not, we used RiboTag+/+;Advillin-Cre+ mice, expressing HA-tagged RPL22 specifically in sensory neurons, to profile their ribosome binding RNAs in nerve injured or uninjured dorsal root ganglion(DRG) tissues using HA-IP sequencing. We analyzed the results together with total RNA transcriptomics conducted under the same conditions. Through this analysis, we found mRNA of Gpr151 gene, one of the transcript that highly responds to injury but does not direct to the ribosomes, promoting axon regeneration by a non-coding RNA function. Overall design: Wild type or RiboTag+/+;Advillin-Cre+ mice were subjected to sciatic nerve injury using forceps in order to profile the injury-responsive changes in total RNA or ribosomal binding RNA transcriptomics within neuronal cell bodies at DRGs. The DRG tissues were collected 72 hours after the injury, and RNAs were extracted from either the whole lysates or the anti-HA immunoprecipitated lysates.