Transcriptional regulatory network during axonal regeneration of dorsal root ganglion neurons:laser-capture microdissection and deep sequencing

作     者：Li-Li Zhao Tao Zhang Wei-Xiao Huang Ting-Ting Guo Xiao-Song Gu Li-Li Zhao;Tao Zhang;Wei-Xiao Huang;Ting-Ting Guo;Xiao-Song Gu

作者机构：Model Animal Research Center and MOE Key Laboratory of Animal Models of DiseaseNanjing UniversityNanjingJiangsu ProvinceChina Key Laboratory of Neuroregeneration of Jiangsu and Ministry of EducationCo-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina School of Medicine and Life SciencesNanjing University of Chinese MedicineNanjingJiangsu ProvinceChina 

出 版 物：《Neural Regeneration Research》 (中国神经再生研究（英文版）)

年 卷 期：2023年第18卷第9期

页      面：2056-2066页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 1001[医学-基础医学(可授医学、理学学位)] 071006[理学-神经生物学] 

基　　金：supported by the National Natural Science Foundation of China,Nos. 31730031 and 32130060 the National Major Project of Research and Development,No. 2017YFA0104700 the Natural Science Foundation of Jiangsu Province,No. BK20202013 (all to XSG)。 

主　　题：Arid5a Atf3 Crem dorsal root ganglion Fosl1 Klf6 laser-capture microdissection neuron smart-seq2 gene expression profile transcription factor 

摘      要：The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified.In high-throughput sequencing,various factors influence the final sequencing results,including the number and size of cells,the depth of sequencing,and the method of cell separation.There is still a lack of research on the detailed molecular expression profile during the regeneration of dorsal root ganglion neuron axon.In this study,we performed lase r-capture microdissection coupled with RNA sequencing on dorsal root ganglion neurons at 0,3,6,and 12 hours and 1,3,and 7 days after sciatic nerve crush in rats.We identified three stages after dorsal root ganglion injury:early(3-12 hours),pre-regeneration(1 day),and regeneration(3-7 days).Gene expression patterns and related function enrichment res ults showed that one module of genes was highly related to axonal regeneration.We verified the up-regulation of activating transcription factor 3(Atf3),Kruppel like factor 6(Klf6),AT-rich inte raction domain 5A(Arid5α),CAMP responsive element modulator(Crem),and FOS like 1,AP-1 transcription factor Subunit(Fosl1) in dorsal root ganglion neurons after injury.Suppressing these transcription factors(Crem,Arid5o,Fosl1 and Klf6) reduced axonal regrowth in vitro.As the hub transcription factor,Atf3 showed higher expression and activity at the preregeneration and regeneration stages.G protein-coupled estrogen receptor 1(Gper1),inte rleukin 12a(Il12α),estrogen receptor 1(ESR1),and interleukin 6(IL6) may be upstream factors that trigger the activation of Atf3 during the repair of axon injury in the early stage.Our study presents the detailed molecular expression profile during axonal regeneration of dorsal root ganglion neurons after peripheral nerve injury.These findings may provide reference for the clinical screening of molecular targets for the treatment of peripheral nerve injury.