Cao Catherine, Nipu Niepukolie, Johnston William, Adil Sally, Wei Lai, Mennigen Jan A
Department of Biology, University of Ottawa, K1 N6 N5 20 Marie Curie, Ottawa, ON, Canada.
Fish Physiol Biochem. 2025 May 7;51(3):92. doi: 10.1007/s10695-025-01505-y.
Several species evolved mechanisms to tolerate periods of severe environmental hypoxia and anoxia. Among them, goldfish are unique as they do not enter a comatose state under such conditions. Taking advantage of the recently published and annotated goldfish genome, we had previously profiled the transcriptomic response of the goldfish brain under normoxic (21 kPa oxygen saturation, N) and hypoxic conditions (2.1 kPa oxygen saturation) after 1 and 4 weeks (1WH, 4WH). Using the RNA-Seq data, we report the occurrence of alternative mRNA splicing (skipped exon, retained intron, alternative 3' or 5' splice sites, and mutually exclusive exons). At 1WH/N, 1004 significant alternative splicing events on 769 gene loci were identified, increasing to 1187 on 963 loci at 4WH/N. There were 305 loci with alternatively spliced transcripts common to both 1WH/N and 4WH/N, 221 of which exhibited the same precise location and splicing mechanism. Specific gene transcripts affected by alternative splicing events were almost entirely different from previously identified differentially expressed genes under chronic hypoxia. GO-term enrichment analyses of gene loci of alternatively spliced transcripts, however, did include similar pathways as previously identified for DEGs. These include epigenetic machinery, ion channel activity (1WH/N), glutamate signaling (4WH/N), endothelial cell function, and ATP hydrolyzation pathways (1WH/N + 4WH/N). We describe selected examples of alternatively spliced transcripts to discuss possible functional relevance in the goldfish brain response to chronic hypoxia. Together, our data identified an additional layer of regulation in brain pathways relevant to hypoxia tolerance in goldfish, which complement previously reported gene expression changes.
几种物种进化出了耐受严重环境低氧和缺氧时期的机制。其中,金鱼很独特,因为它们在这种条件下不会进入昏迷状态。利用最近公布并注释的金鱼基因组,我们之前已分析了金鱼大脑在常氧(氧饱和度21 kPa,N)和低氧条件(氧饱和度2.1 kPa)下1周和4周后(1WH,4WH)的转录组反应。利用RNA测序数据,我们报告了可变mRNA剪接(外显子跳跃、内含子保留、可变3'或5'剪接位点以及互斥外显子)的发生情况。在1WH/N时,在769个基因位点上鉴定出1004个显著的可变剪接事件,在4WH/N时增加到963个位点上的1187个。在1WH/N和4WH/N中共有305个位点具有可变剪接转录本,其中221个表现出相同的精确位置和剪接机制。受可变剪接事件影响的特定基因转录本几乎与之前鉴定出的慢性低氧条件下的差异表达基因完全不同。然而,可变剪接转录本基因位点的GO术语富集分析确实包括了与之前鉴定出的差异表达基因相似的途径。这些途径包括表观遗传机制、离子通道活性(1WH/N)、谷氨酸信号传导(4WH/N)、内皮细胞功能以及ATP水解途径(1WH/N + 4WH/N)。我们描述了可变剪接转录本的选定示例,以讨论其在金鱼大脑对慢性低氧反应中的可能功能相关性。总之,我们的数据确定了金鱼与低氧耐受性相关的大脑途径中的另一层调控,这补充了之前报道的基因表达变化。