Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, 300071, PR China.
Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, 300071, PR China.
Dev Comp Immunol. 2020 May;106:103630. doi: 10.1016/j.dci.2020.103630. Epub 2020 Jan 22.
Innate immunity plays an important role in invertebrates because it provides the first line of protection by recognizing invading microbial pathogens and then activating downstream signaling pathways. However, until now, increasing reports of clam diseases did not include those of Anadara sativa, which are widely distributed and economically important maritime clams. In the present study, transcriptome libraries of untreated (termed H) and Vibrio anguillarum-challenged (termed HV) A. sativa hepatopancreases were constructed and sequenced using the Illumina HiSeq4000 platform. In total, we obtained 78,012,510 and 84,937,516 clean reads from 80,006,030 to 86,871,742 raw data reads, respectively, assembled by different software programs. Furthermore, 150,274 unigenes were generated from 196,003 transcripts, with an N50 length of 1088 bp, and then annotated with the SwissProt, NR, NT, PFAM, KO, GO, KOG and KEGG databases. Moreover, 3982 differentially expressed unigenes (H vs HV) were determined, with 3583 upregulated and 399 downregulated genes. Among these differentially expressed unigenes, 207 unigenes were found using KEGG annotation in 16 immune-related signaling pathways, such as Toll-like receptor (TLR), NOD-like receptor (NLR), and RIG-I-like receptor (RLR) signaling pathways. Finally, we selected 11 full-length TLRs and classified them into 3 groups, namely, one V-TLR, four Ls-TLR and six sP-TLR; furthermore, we validated the increased expression patterns of the 11 TLRs in response to LPS injection. In summary, these results revealed multiple findings on potential immune-related genes, such as the differential expression analysis and annotation based on the A. sativa transcriptome in response to V. anguillarum stimulation, and explored the molecular and expression characterization of A. sativa TLRs, which provide new insights into the innate immune responses and defense mechanisms in shellfish.
先天免疫在无脊椎动物中起着重要作用，因为它通过识别入侵的微生物病原体，然后激活下游信号通路，提供第一道保护。然而，直到现在，越来越多的贝类疾病报告都没有包括广泛分布且具有经济重要性的海洋蛤类——缢蛏。在本研究中，构建了未经处理（命名为 H）和鳗弧菌刺激（命名为 HV）缢蛏肝胰腺的转录组文库，并使用 Illumina HiSeq4000 平台进行测序。总共从 80006030 到 86871742 个原始数据读取中获得了 78012510 和 84937516 个清洁读取，分别通过不同的软件程序进行组装。此外，从 196003 个转录本中生成了 150274 个基因，N50 长度为 1088bp，然后在 SwissProt、NR、NT、PFAM、KO、GO、KOG 和 KEGG 数据库中进行注释。此外，确定了 3982 个差异表达的基因（H 与 HV），其中 3583 个上调，399 个下调。在这些差异表达的基因中，在 16 个免疫相关信号通路中，使用 KEGG 注释发现了 207 个基因，如 Toll 样受体（TLR）、NOD 样受体（NLR）和 RIG-I 样受体（RLR）信号通路。最后，我们选择了 11 个全长 TLR 并将它们分为 3 组，即 1 个 V-TLR、4 个 Ls-TLR 和 6 个 sP-TLR；此外，我们验证了 LPS 注射后 11 个 TLR 的表达模式增加。总之，这些结果揭示了关于潜在免疫相关基因的多个发现，例如基于缢蛏转录组对鳗弧菌刺激的差异表达分析和注释，以及探索缢蛏 TLR 的分子和表达特征，为贝类先天免疫反应和防御机制提供了新的见解。
