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缺氧再复氧应激调节中华绒螯蟹肝胰腺转录组。

Hypoxia-reoxygenation stress modulates the hepatopancreas transcriptome of Chinese mitten crab Eriocheir sinensis.

机构信息

Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing 210017, China.

Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing 210017, China; College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

出版信息

Gene. 2021 Mar 1;771:145361. doi: 10.1016/j.gene.2020.145361. Epub 2020 Dec 16.

Abstract

Hypoxia is a critical, but frequently overlooked problem, which commonly exists in Chinese mitten crab rearing. However, little information is available on the molecular mechanisms of the detrimental effects of hypoxia in this species. In the present study, crabs were subjected to acute hypoxia stress (DO 1.0 mg/L), followed by reoxygenation (DO 6.8 mg/L). Hepatopancreas from five groups of crabs (three to four crabs per group), including normoxia, hypoxia for one and six hours, and reoxygenation for one and 12 h, were used for transcriptome sequencing. The pooled total RNA of all samples were utilized to reconstruct a reference transcriptome with PacBio RS II sequencing, obtaining 49.19 G clean data, with a mean length of 2,180 bp. Seventeen cDNA libraries were constructed and sequenced to identify differentially expressed genes (DEGs) among the different samples (FDR < 0.05 and |log2 fold change| ≥1). A total of 103 and 251 DEGs were identified when exposed to hypoxia for one and six hours, respectively. Totally 462 and 673 DEGs were identified during reoxygenation at 1 and 12 h, respectively. Among these DEGs, two transcripts with complete ORFs were identified to encode hypoxia-inducible factor 1 (Es-Hif-1α/β), which is a transcriptional activator of various genes correlated to the cellular adaptive responses to hypoxia. Es-Hif-1a/β expressions were significantly upregulated when exposed to hypoxia treatment, and no significant difference was observed for Es-Hif-1α between hypoxic treatment for 6 h and reoxygenation. The significant KEGG enrichment revealed that the DEGs under hypoxia were mainly enriched in "PPAR signaling pathway", "Gap junction" and "Phototransduction-fly". Compared with crabs in normoxia, even with 12 h of reoxygenation, the hepatopancreas transcriptome under hypoxia remained severely affected, implying its adverse effect on growth and development, or increased susceptibility to diseases.

摘要

缺氧是一个关键但经常被忽视的问题,在中国绒螯蟹养殖中普遍存在。然而,关于这种物种缺氧的有害影响的分子机制的信息很少。在本研究中,将蟹置于急性缺氧应激(DO 1.0 mg/L)下,然后再进行复氧(DO 6.8 mg/L)。来自五组蟹(每组三到四只蟹)的肝胰腺,包括正常氧、缺氧 1 小时和 6 小时以及复氧 1 小时和 12 小时,用于转录组测序。所有样本的总 RNA 用于 PacBio RS II 测序构建参考转录组,获得 49.19 G 清洁数据,平均长度为 2180 bp。构建了 17 个 cDNA 文库并进行测序,以鉴定不同样本之间的差异表达基因(FDR < 0.05 和 |log2 倍变化|≥1)。分别在缺氧 1 小时和 6 小时时鉴定到 103 和 251 个差异表达基因。在复氧 1 小时和 12 小时时分别鉴定到 462 和 673 个差异表达基因。在这些差异表达基因中,鉴定到两个具有完整 ORF 的转录本,分别编码缺氧诱导因子 1(Es-Hif-1α/β),它是与细胞对缺氧的适应性反应相关的各种基因的转录激活因子。当暴露于缺氧处理时,Es-Hif-1α/β 的表达明显上调,而在 6 小时缺氧处理和复氧之间,Es-Hif-1α 没有显著差异。显著的 KEGG 富集表明,缺氧下的差异表达基因主要富集在“PPAR 信号通路”、“间隙连接”和“光转导-蝇”中。与正常氧下的蟹相比,即使复氧 12 小时,缺氧下的肝胰腺转录组仍受到严重影响,这意味着其对生长和发育有不利影响,或者增加了患病的易感性。

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