RNA sequencing of Dorsal Root Ganglia upon axonal injury, in presence or absence of NAC (N-acetyl cysteine), and upon H2O2 nerve treatment

Purpose: A) compare the response to an axonal injury in the peripheral vs central nervous system B) clarify the mechanism of Reactive Oxygen Species-dependent axonal regeneration Results: we found that after peripheral axonal injury there is a higher number of differentially expressed genes with respect to central injury. Moreover,inflammation related genes and regeneration-associated genes were differentially regulated after peripheral injury and H2O2, but less after central injury. Signalling pathways and biological processes related to injury-dependent nerve regeneration were reduced with the addition of NAC at the time of sciatic injury compared to vehicle. Method: RNA-seq from sciatic L4-L6 DRG tissue 24 hours after the following treatments: a) spinal cord dorsal column injury b) laminectomy c) sciatic nerve injury in presence of saline d) sham in presence of saline, e) sciatic nerve injury in presence of 30 mM NAC (N-acetyl-cysteine), f) sham in presence of 13 mM H2O2. Pool of sciatic DRGs from 2 mice were used for each sample. DRG were collected in RNase later reagent, and crushed with RNase free micropestle for RNA extraction.