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C57BL/6J 小鼠角膜中 mRNA 表达的日常节律转录组学分析。

Transcriptional Profiling of Daily Patterns of mRNA Expression in the C57BL/6J Mouse Cornea.

机构信息

Henan Provincial People's Hospital and People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital , Zhengzhou , China.

International Ocular Surface Research Center and Institute of Ophthalmology, Key Laboratory for Regenerative Medicine, Jinan University Medical School , Guangzhou , China.

出版信息

Curr Eye Res. 2019 Oct;44(10):1054-1066. doi: 10.1080/02713683.2019.1625408. Epub 2019 Jun 25.

Abstract

: The purpose of this study was to determine how the transcriptome of the murine cornea adapts to diurnal changes in physiology. : C57BL/6J mice were maintained under a 12-h light/12-h dark (LD) cycle for two weeks. Corneas were collected from euthanized mice at Zeitgeber time (ZT) 0, 3, 6, 9, 12, 15, 18, and 21. Total RNA was extracted and subjected to RNA sequencing (RNA-Seq). A JTK_CYCLE algoithm and other software tools were used to analyze the transcriptional data to determine the periodicity, rhythmicity, and amplitude of the transcripts. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the enrichment of cycling transcripts. : Approximately 24% of the total transcripts from the murine corneal genome were rhythmically expressed over an LD cycle. GO analysis showed that these cycling genes are primarily involved in cellular and metabolic processes. A KEGG pathway analysis identified 6 branches and 44 pathways that encode the gene outputs necessary for basic cellular functions and processes. More importantly, most of the rhythmic genes between the day and night are enriched in their own unique pathways in addition to some common pathways. Furthermore, most of the rhythmic gene expression was concentrated in the 12-h and 24-h periods. A comparative analysis of GO and KEGG showed large differences in metabolic processes, but not cellular processes. Finally, the murine cornea also rhythmically expressed 11 canonical components of circadian clock genes over an LD cycle at the transcriptional level. : One fourth of the corneal transcriptome follows a rhythmic expression pattern involved in basic molecular and cellular mechanisms. This implies that the time of day contributes significantly to the overall temporal organization of the corneal transcriptome.

摘要

本研究旨在确定小鼠角膜的转录组如何适应生理昼夜变化。将 C57BL/6J 小鼠维持在 12 小时光照/12 小时黑暗(LD)周期下两周。在 Zeitgeber 时间(ZT)0、3、6、9、12、15、18 和 21 时从安乐处死的小鼠中采集角膜。提取总 RNA 并进行 RNA 测序(RNA-Seq)。使用 JTK_CYCLE 算法和其他软件工具分析转录数据,以确定转录本的周期性、节律性和振幅。使用基因本体论(GO)和京都基因与基因组百科全书(KEGG)分析循环转录本的富集情况。

在 LD 周期中,大约 24%的小鼠角膜基因组总转录物呈节律性表达。GO 分析表明,这些循环基因主要参与细胞和代谢过程。KEGG 途径分析确定了 6 个分支和 44 条途径,这些途径编码了基本细胞功能和过程所需的基因产物。更重要的是,白天和黑夜之间的大多数节律基因除了一些共同途径外,还富集在自己独特的途径中。此外,大多数节律基因的表达主要集中在 12 小时和 24 小时周期内。GO 和 KEGG 的比较分析表明,代谢过程存在很大差异,但细胞过程没有。最后,在 LD 周期内,小鼠角膜在转录水平上也节律性地表达了 11 个生物钟基因的典型组成部分。

角膜转录组的四分之一遵循涉及基本分子和细胞机制的节律表达模式。这意味着一天中的时间对角膜转录组的整体时间组织有重要贡献。

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