Abstract
Peripheral nerve injury could lead to chronic neuropathic pain. Understanding transcriptional changes induced by nerve injury could provide fundamental insights into the complex pathogenesis of neuropathic pain. Gene expression profiles of dorsal root ganglia (DRG) in neuropathic pain condition have been studied. However, little is known about transcriptomic changes in individual DRG neurons after peripheral nerve injury. Here we performed single-cell RNA sequencing on dissociated mouse DRG cells after spared nerve injury (SNI). In addition to DRG neuron types that are found under physiological conditions, we identified three SNI-induced neuronal clusters (SNIICs) characterized by the expression of Atf3/Gfra3/Gal (SNIIC1), Atf3/Mrgprd (SNIIC2) and Atf3/S100b/Gal (SNIIC3). These SNIICs originated from Cldn9+/Gal+, Mrgprd+ and Trappc3l+ DRG neurons, respectively. Interestingly, SNIIC2 switched to SNIIC1 by increasing Gal and reducing Mrgprd expression 2 days after nerve injury. Inferring the gene regulatory networks after nerve injury, we revealed that activated transcription factors Atf3 and Egr1 in SNIICs could enhance Gal expression while activated Cpeb1 in SNIIC2 might suppress Mrgprd expression within 2 days after SNI. Furthermore, we mined the transcriptomic changes in the development of neuropathic pain to identify potential analgesic targets. We revealed that cardiotrophin-like cytokine factor 1, which activates astrocytes in the dorsal horn of spinal cord, was upregulated in SNIIC1 neurons and contributed to SNI-induced mechanical allodynia. Therefore, our results provide a new landscape to understand the dynamic course of neuron type changes and their underlying molecular mechanisms during the development of neuropathic pain.
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Data availability
Most datasets generated in the current study are available in the GSE155622. The other datasets analyzed in the current study are included in Li et al.14 Processed data also available in zhanglab.shbrainproject.com: 100. R scripts are available from the corresponding author upon reasonable request.
Change history
19 May 2021
A Correction to this paper has been published: https://doi.org/10.1038/s41422-021-00503-y
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Acknowledgements
We thank Dr. Ping Hu for the comments and suggestions on the experimental design for the transcriptional regulation section. This work was supported by National Natural Science Foundation of China (31671094, 31630033, and 31600853), Chinese Academy of Sciences (QYZDYSSW-SMC007), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB39050100), the International Partnership Program of Chinese Academy of Sciences (172644KYS820170004), Science and Technology Commission of Shanghai Municipality (18JC1420301) and Innovation Fund for Medical Sciences of Chinese Academy of Medical Sciences (2019-I2M-5-082).
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X.Z., C.L., L.B., and L.W. conceived and designed the project. S.W. and D.W. performed 10× scRNA-seq. Y.C. performed single-cell experiments by Smart-seq2. K.W. and C.L. analyzed scRNA-seq data. S.W., K.W., and Y.L. performed histological experiments. S.W. performed behavioral experiments. K.W., C.L., and X.Z. wrote the manuscript.
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Wang, K., Wang, S., Chen, Y. et al. Single-cell transcriptomic analysis of somatosensory neurons uncovers temporal development of neuropathic pain. Cell Res 31, 904–918 (2021). https://doi.org/10.1038/s41422-021-00479-9
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DOI: https://doi.org/10.1038/s41422-021-00479-9
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