Unveiling the Time Course of Neuropathic Pain via Single-Cell Analysis
Summary
Chronic neuropathic pain is a common complication of peripheral nerve injury. In order to gain a better understanding of the transcriptional changes resulting from nerve injury, researchers conducted single-cell RNA sequencing on dissociated mouse dorsal root ganglia (DRG) cells after spared nerve injury (SNI). The study revealed three SNI-induced neuronal clusters (SNIICs) that originated from distinct DRG neurons, in addition to those identified under physiological conditions. By inferring gene regulatory networks after nerve injury, they found that activated transcription factors Atf3 and Egr1 in SNIICs could enhance Gal expression, while Cpeb1 in SNIIC2 might suppress Mrgprd expression within 2 days after SNI. The study also identified a potential analgesic target for SNI-induced mechanical allodynia.
Research Criteria
They used scRNA-seq (single cell RNA sequencing) to examine the single-cell transcriptomes of mouse DRG at various time points after SNI, a widely used neuropathic pain model.
Fig.1 Experimental design. (Wang, 2021)
Sample Type
Mouse DRG tissues (after SNI 6h, 24h, 2d, 7d, 14d)
Result—Cell Heterogeneity in DRG
The authors aimed to decipher the single-cell transcriptomes of adult mouse DRG neurons under neuropathic pain conditions. The rodents underwent unilateral SNI, a model that elicits robust and prolonged mechanical allodynia. They subsequently performed scRNA-seq on the cell dissociates from L4 and L5 DRGs on the side of the SNI at different time points. After stringent filtering, including the removal of empty droplets, outliers, cell debris, and inferred doublets, they retained 36,810 cells, including 6,935 neurons. They identified nine major DRG cell types with distinct molecular markers, including primary sensory neurons, SGCs, Schwann cells, VECs, VSMCs, VECCs, fibroblasts, immune cells, and RBCs. The non-neuron cell classification after SNI was consistent with that under normal conditions, and the authors have provided a comprehensive gene expression profile for each DRG cell type.
Fig.2 The heterogeneity of DRG cells in the SNI model. (Wang, 2021)
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Reference
- Wang, K.L.; et al. Single-cell transcriptomic analysis of somatosensory neurons uncovers temporal development of neuropathic pain. Cell Research. 2021, 31: 904-918.
