Sex-specific transcriptome of spinal microglia in neuropathic pain due to peripheral nerve injury

Neuropathic pain is a prevalent and debilitating chronic disease that is characterized by activation in glial cells in various pain-related regions within the central nervous system. Recent studies have suggested a sexually dimorphic role of microglia in the maintenance of neuropathic pain in rodents. Here, we utilized RNA sequencing analysis of microglia to identify whether there is a common neuropathic microglial signature and characterize the sex differences in microglia in pain-related regions in nerve injury and chemotherapy-induced peripheral neuropathy mouse models. Whilst mechanical allodynia and behavioral changes were observed in all models, transcriptomic analysis of microglia revealed no common transcriptional changes in spinal and supraspinal regions and in different neuropathic models. However, there was a substantial change in microglial gene expression within the ipsilateral lumbar spinal cord 7-days after chronic constriction injury (CCI) of the sciatic nerve. Both sexes upregulated genes associated with inflammation, phagosome, and lysosome activation, though males revealed a prominent global transcriptional shift not observed in female mice. This study demonstrates a lack of a common neuropathic microglial signature and indicates distinct sex differences in spinal microglia, suggesting they contribute to the sex-specific pain processing following nerve injury. Overall design: RNA-seq of isolated microglia after chronic constriction injury or sham surgery (day 7) from male and female mice. Four blocks of CNS were run following CCI or Sham surgery ; (1) the ipsilateral (left) lumbar spinal cord (L3-L5); (2) the medial prefrontal cortex, hippocampus and amygdala; (3) posterior thalamus and S1 cortex relating to the hindlimb; and (4) the periaqueductal gray and rostroventral medulla.