The Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao, 266003, PR China.
The Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao, 266003, PR China.
Fish Shellfish Immunol. 2019 Nov;94:497-509. doi: 10.1016/j.fsi.2019.09.043. Epub 2019 Sep 18.
As a crucial neuroendocrine-immune factor, dopamine (DA) could regulate the immune system of Litopenaeus vannamei. To understand the immune mechanisms and regulatory pathways of DA in L. vannamei, the transcriptome analysis of hemocytes of L. vannamei with injection of DA (10 mol/shrimp) at 3 and 12 h were performed in this study. Moreover, quantitative real-time PCR (qPCR) method was applied to validate the accuracy of transcriptome sequencing and analyze the expression pattern of candidate differentially expressed genes (DEGs) at different time points (0, 3, 6, 12, and 24 h) after DA injection. The results showed that a total of 51382 unigenes with a N50 length of 2341 bp were generated. And 1397 and 457 DEGs were obtained by comparative transcriptome at 3 and 12h respectively. Moreover, the results of functional annotation and enriched pathway showed that the DEGs were involved in phagosome (ko04145), lysosome (ko04142), Endocytosis (ko04144), and NOD-like receptor signaling pathway (ko04621). Besides, the Pearson's correlation coefficient (R) between transcriptome sequencing and qPCR was 0.845, which confirmed the reliability of the transcriptome sequencing results and the accuracy of assembly. Furthermore, the expression pattern of 15 candidate DEGs, containing 9 up-regulated and 6 down-regulated DEGs at 3 h, indicated the regulation of DA in physiological functions especially in the immune system. Therefore, these results revealed that DA induced the expressions of membrane receptors or proteins, activated intracellular signaling pathways, regulated cellular and humoral immune systems, controlled antioxidation and apoptosis, and was involved in the regulation of neuroendocrine system. These findings are helpful to promote the understanding on the effects of biogenic amines on physiological functions and regulatory networks of crustacean, and offer a substantial material and foundation for researching the immune response of crustacean.
作为一种重要的神经内分泌免疫因子，多巴胺（DA）可以调节凡纳滨对虾的免疫系统。为了了解 DA 在凡纳滨对虾中的免疫机制和调控途径，本研究对注射 DA（10mol/虾）后 3 和 12h 的凡纳滨对虾血细胞进行了转录组分析。此外，还应用实时荧光定量 PCR（qPCR）方法验证了转录组测序的准确性，并分析了 DA 注射后不同时间点（0、3、6、12 和 24h）候选差异表达基因（DEGs）的表达模式。结果显示，共生成了 51382 条长度为 2341bp 的 unigenes。通过比较转录组，在 3h 和 12h 时分别获得了 1397 个和 457 个 DEGs。此外，功能注释和富集途径的结果表明，DEGs 参与了吞噬体（ko04145）、溶酶体（ko04142）、内吞作用（ko04144）和 NOD 样受体信号通路（ko04621）。此外，转录组测序和 qPCR 的 Pearson 相关系数（R）为 0.845，这证实了转录组测序结果的可靠性和组装的准确性。此外，15 个候选 DEGs 的表达模式，包括 3h 时 9 个上调和 6 个下调的 DEGs，表明了 DA 对生理功能，尤其是免疫系统的调节作用。因此，这些结果揭示了 DA 诱导了膜受体或蛋白的表达，激活了细胞内信号通路，调节了细胞和体液免疫系统，控制了抗氧化和细胞凋亡，并参与了神经内分泌系统的调节。这些发现有助于促进对生物胺对甲壳动物生理功能和调控网络影响的理解，为研究甲壳动物的免疫反应提供了重要的物质和基础。
