FACS purified Nav1.8 lineage sensory neuron whole transcriptome changes following peripheral inflammation (CFA) or sciatic nerve injury (SNI).

The goal of this study was to analyze global gene expression in FACS purified Nav1.8 lineage sensory neurons, which include nociceptor neurons that detect damaging/noxious stimuli, following peripheral inflammation by intraplantar injection of Complete Freund's Adjuvant (CFA) or Sciatic Nerve Injury (SNI) by nerve transection. Nav1.8 Trangsgenic TdTomato+ neurons were purified from Lumbar L4-L6 dorsal root ganglia (DRG) by flow cytometry from mice on the ipsilateral or contralateral sides, following Complete Freund's Adjuvant injection (day 1) or sciatic nerve transection (day 5). Neurons were then analyzed for transcriptional gene expression by microarray analysis. Nav1.8-Cre transgenic mice (also known as SNS-Cre mice) were bred with Rosa26-TdTomato mice (ai14 mice from jackson Labs) to generate Nav1.8-Cre Tg+ TdTomato+ reporter progeny, where peripheral sensory neurons expresing the sodium channel Nav1.8 are genetically marked with red fluroescence. In one group of Nav1.8-Cre Tg/TdTomato mice, mice were injected intraplantarly with complete freunds adjuvant. In another group of mice, mice underwent complete sciatic nerve transection. Lumbar region dorsal root ganglia (DRG) were dissected from contralateral or ipsilateral sides of CFA injected mice or sciatic nerve transected mice (3 mice were pooled/sample). Using flow cytometry, highly red fluorescent neurons were Facs purified, total RNA was extracted, and processed for microarray analysis. For additional comparisons, Nav1.8-Cre Tg/TdTomato lumbar neurons were previously deposited at GEO database (GSE46546).