Deep RNA-seq of male and female murine sensory neuron subtypes after nerve injury

Supplemental Digital Content is Available in the Text.Using sex, injury, and time factors, RNA-seq was performed on dorsal root ganglia subtypes isolated through fluorescent activated cell sorting. This was analyzed and curated as a resource for others. Dorsal root ganglia (DRG) neurons have been well described for their role in driving both acute and chronic pain. Although nerve injury is known to cause transcriptional dysregulation, how this differs across neuronal subtypes and the impact of sex is unclear. Here, we study the deep transcriptional profiles of multiple murine DRG populations in early and late pain states while considering sex. We have exploited currently available transgenics to label numerous subpopulations for fluorescent-activated cell sorting and subsequent transcriptomic analysis. Using bulk tissue samples, we are able to circumvent the issues of low transcript coverage and drop-outs seen with single-cell data sets. This increases our power to detect novel and even subtle changes in gene expression within neuronal subtypes and discuss sexual dimorphism at the neuronal subtype level. We have curated this resource into an accessible database for other researchers (https://livedataoxford.shinyapps.io/drg-directory/). We see both stereotyped and unique subtype signatures in injured states after nerve injury at both an early and late timepoint. Although all populations contribute to a general injury signature, subtype enrichment changes can also be seen. Within populations, there is not a strong intersection of sex and injury, but previously unknown sex differences in naive states-particularly in A & beta;-RA + A & delta;-low threshold mechanoreceptors-still contribute to differences in injured neurons.

Automated summary

This study performed RNA-seq analysis on subtypes of dorsal root ganglia (DRG) neurons to investigate their transcriptional profiles in acute and chronic pain states, while considering sex differences. The results revealed both stereotyped and unique subtype signatures in injured states and previously unknown sex differences in naive states, contributing to differences in injured neurons.