近年来,频繁的急性温度变化对鱼类的生理和生存构成了严重威胁。本研究利用RNA-Seq技术分析了在各种急性温度条件下(16°C,200.50C,24ØC,28□C和32□C)。通过综合分析,我们确定了111509个差异表达基因(DEGs),随着温度升高而显著上调的基因集(谱19),和两个与急性温度变化显着相关的加权基因共表达网络分析(WGCNA)模块。此外,我们确定了28个转录因子,在急性温度变化下的氧化应激和能量代谢中起关键作用。我们的研究结果表明,与对照组(24°C)相比,KEGG功能富集分析显示DEGs在细胞周期中显著富集,DNA复制,和p53信号通路,具有抑制表达的总体趋势。这表明,保持细胞稳定性和减少细胞损伤是温和草应对急性温度变化的有效适应机制。通过STEM分析和与高温应力相关的黑色WGCNA模块,我们发现了与能量代谢相关的通路和中枢基因的显著上调,包括氧化磷酸化,TCA循环,嘌呤代谢,和谷胱甘肽代谢,以及MAPK信号通路和AMPK信号通路等信号转导通路的核心作用,协同调节能源生产。在急性低温胁迫下,绿松石WGCNA模块强调了与蛋白质合成和加工相关的核糖体和剪接体途径相关的hub基因的显着上调,以及钙信号通路的激活,在低温适应过程中对维持细胞功能起着重要作用。这些发现为热鱼适应快速温度变化提供了关键的理论和分子基础。
In recent years, frequent acute temperature changes have posed a serious threat to the physiology and survival of fish. This study utilized RNA-Seq technology to analyze the transcriptional dynamics in the muscle tissues of Acrossocheilus wenchowensis under various acute temperature conditions (16◦C, 20◦C, 24◦C, 28◦C and 32◦C). Through comprehensive analysis, we identified 11509 differentially expressed genes (DEGs), a gene set (profiles 19) that was significantly up-regulated with increasing temperature, and two weighted gene co-expression network analysis (WGCNA) modules that were significantly correlated with acute temperature changes. Furthermore, we identified 28 transcription factors that are pivotal in oxidative stress and energy metabolism under acute temperature changes. Our results showed that, compared to the control group (24°C), KEGG functional enrichment analysis revealed significant enrichment of DEGs in the cell cycle, DNA replication, and p53 signaling pathway, with an overall trend of suppressed expression. This indicates that maintaining cell stability and reducing cell damage is an effective adaptive mechanism for A. wenchowensis to cope with acute temperature changes. Through STEM analysis and the black WGCNA module associated with high-temperature stress, we identified significant up-regulation of pathways and hub genes related to energy metabolism including oxidative phosphorylation, TCA cycle, purine metabolism, and glutathione metabolism, as well as the central roles of signal transduction pathways such as MAPK signaling pathway and AMPK signaling pathway, which synergistically regulate energy production. Under acute low-temperature stress, the turquoise WGCNA module highlighted significant up-regulation of hub genes associated with Ribosomal and Spliceosomal pathways related to protein synthesis and processing, as well as activation of calcium signaling pathways, which plays an important role in maintaining cellular function during low-temperature adaptation. These findings provide a critical theoretical and molecular basis for the adaptation of eurythermal fish to rapid temperature changes.