College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China; College of Marine Science and Engineering, Nanjing Normal University, Nanjing, 210046, China.
Jiangsu Key Laboratory for Biofunctional Molecules, College of Life Science and Chemistry, Jiangsu Second Normal University, Nanjing, 210013, China.
Fish Shellfish Immunol. 2018 Dec;83:115-122. doi: 10.1016/j.fsi.2018.09.005. Epub 2018 Sep 6.
The immune deficiency (IMD) signal pathway mediates innate immunity against Gram-negative bacteria in crustaceans. In the present study, an IMD homolog (MnIMD) from the oriental river prawn, Macrobrachium nipponense was identified. The full-length cDNA of MnIMD was 782bp with an open reading frame of 549 bp that encodes a putative protein of 182 amino acids including a death domain at the C-terminus. Multiple alignment analysis showed that IMDs in prawn M. nipponense and other crustaceans shared high similarity. The recombinant protein of MnIMD was expressed and purified for further functional analyses. Western blot analysis indicated that MnIMD was present in many tissues, but with the highest level in the gills, which was consistent with the qRT-PCR results. After Vibrio parahaemolyticus challenge, MnIMD was significantly induced in gills. RNA interference analysis showed that the IMD pathway was involved in regulating the expression of different antimicrobial peptide (AMP) genes, including Cru4 and Cru6. These results are helpful in promoting research on the innate immunity in M. nipponense.
免疫缺陷(IMD)信号通路介导甲壳动物对革兰氏阴性菌的先天免疫。本研究从日本沼虾(Macrobrachium nipponense)中鉴定出一个 IMD 同源物(MnIMD)。MnIMD 的全长 cDNA 为 782bp,开放阅读框为 549bp,编码一个推测的 182 个氨基酸的蛋白质,其 C 末端有一个死亡结构域。多重比对分析表明,MnIMD 与其他甲壳动物的 IMDs 具有高度相似性。重组 MnIMD 蛋白得到表达和纯化,用于进一步的功能分析。Western blot 分析表明,MnIMD 存在于许多组织中,但在鳃中表达水平最高,与 qRT-PCR 结果一致。在副溶血弧菌攻毒后,MnIMD 在鳃中显著诱导。RNA 干扰分析表明,IMD 途径参与调节不同抗菌肽(AMP)基因的表达,包括 Cru4 和 Cru6。这些结果有助于促进对日本沼虾先天免疫的研究。