多疣壁虎断尾后脊髓再生过程中mRNA和微小RNA差异表达谱的分析

doi:10.16098/j.issn.0529-1356.2023.01.001

摘要/Abstract

摘要：

目的分析多疣壁虎断尾后脊髓再生过程中相关mRNA和微小RNA（miRNA）的表达变化，探讨差异表达的mRNA和miRNA在脊髓损伤再生过程中的生物学作用。方法构建多疣壁虎断尾模型，50只多疣壁虎分为正常组、断尾15d组和断尾25d组，每组5只实验重复3次，余5只备用。收集各组标本，提取各组RNA 并进行高通量测序。生物信息学分析鉴定组间差异表达的mRNA、miRNA，基因本体论（GO）富集分析差异表达的mRNA功能注释,构建脊髓再生相关的miRNA和mRNA基因调控网络。结果经测序分析多疣壁虎正常和新生脊髓中mRNA、miRNA的差异表达，断尾15d组和断尾25d组分别鉴定到538、510个差异mRNA表达和446、127个差异miRNA表达。GO分析发现，差异表达的mRNA聚集于与细胞增殖、神经发育相关的生物学过程。在脊髓再生相关miRNA及其靶基因调控网络中，断尾15d组有21个mRNA表达下调，被41个上调的miRNA负向调控；12个mRNA表达上调，受到29个下调的miRNA调控。在断尾25d组中，8个mRNA表达下调，被10个上调的miRNA负向调控；20个mRNA表达上调，受到32个下调的miRNA调控。结论通过对多疣壁虎再生脊髓中miRNA、mRNA差异表达分析，初步揭示了mRNA、miRNA在脊髓再生中表达变化的规律，为阐明脊髓再生的分子机制提供了实验数据。

关键词: 脊髓再生, 断尾, 微小RNA, 高通量测序, 多疣壁虎

Abstract:

Objective To analyze the expression changes of related mRNA and microRNA（miRNA）during spinal cord regeneration after tail amputation of Gekko japonicus, and to explore the biological effects of differentially expressed mRNA and miRNA during spinal cord regeneration. Methods Fifty Gekko japonicus, the tail amputation model of Gekko japonicus was constructed, divided into normal group, 15days tail amputation group, and 25days tail amputation group, 5 in each group, repeat the experiment 3 times, 5 spare. Samples of each group were collected, RNA of each group was extracted and high-throughput sequencing. Bioinformatics analysis identifies differentially expressed mRNA and miRNA between groups, Gene Ontology(GO) enrichment analysis of differentially expressed mRNA functional annotations, and construction of miRNA and mRNA gene regulatory networks related to spinal cord regeneration. Results The differential expression of mRNA and miRNA in the normal and newborn spinal cords of Gekko japonicus was analyzed by sequencing. The 15days and 25days tail amputation groups identified 538 and 510 differential mRNA expressions and 446, 127 differential miRNA expressions, respectively. GO analysis found that the differentially expressed mRNA aggregated in biological processes related to cell proliferation and neurodevelopment. In the spinal cord regeneration-related miRNA and its target gene regulatory network, 21 mRNA expression was down-regulated in the 15days tail amputation group, which was regulated negatively by 41 up-regulated miRNAs; 12 mRNA expression was up-regulated and was regulated by 29 down-regulated miRNAs. In the 25days tail amputation group, 8 mRNA expression was down-regulated and regulated negatively by 10 up-regulated miRNAs; 20 mRNA expression was up-regulated and regulated by 32 down-regulated miRNAs. Conclusion Through the analysis of the differential expression of miRNA and mRNA in the regenerated spinal cord of Gekko japonicus, the expression changes of mRNA and miRNA in spinal cord regeneration were initially revealed, which provided experimental data for elucidating the molecular mechanism of spinal cord regeneration.

Key words: Spinal cord regeneration, Tail amputation, MicroRNA, High-throughput sequencing, Gekko japonicus

中图分类号:

Q189

胡晓静张明敏吕广明. 多疣壁虎断尾后脊髓再生过程中mRNA和微小RNA差异表达谱的分析[J]. 解剖学报, 2023, 54(1): 6-12.

HU Xiao-jing ZHANG Ming-min Lü Guang-ming. Preliminary analysis of mRNA and microRNA differential expression profiles in spinal cord regeneration of Gekko japonicus after tail amputation[J]. Acta Anatomica Sinica, 2023, 54(1): 6-12.

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