School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China.
State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
Int J Mol Sci. 2024 Oct 1;25(19):10532. doi: 10.3390/ijms251910532.
Ras-related associated with diabetes (RRAD) is a member of the Ras GTPase superfamily that plays a role in several cellular functions, such as cell proliferation and differentiation. In particular, the superfamily acts as an NF-κB signaling pathway inhibitor and calcium regulator to participate in the immune response pathway. A recent transcriptome study revealed that was expressed in the spleen of disease-resistant Japanese flounder () individuals compared with disease-susceptible individuals, and the results were also verified by qPCR. Thus, the present study aimed to explore how regulates antimicrobial immunity via the NF-κB pathway. First, the coding sequence of was identified. The sequence was 1092 bp in length, encoding 364 amino acids. Based on and structural relationship analyses, appeared to be more closely related to teleosts. Next, expression differences between disease-resistant and disease-susceptible individuals in immune-related tissues were evaluated, and the results revealed that was expressed preferentially in the spleen of disease-resistant individuals. In response to infection, expression in the spleen changed. In vitro, co-culture was carried out to assess the hypo-methylated levels of the promoter in the disease-resistant spleen, which was consistent with the high mRNA expression. The siRNA-mediated knockdown of performed with the gill cell line of affected many -network-related genes, i.e., , , , , , , , , , , , and others, as well as some inflammation-related genes, such as and . In addition, flow cytometry analysis showed that overexpression was more likely to induce cell apoptosis, with establishing a link between 's function and its potential roles in regulating the NF-κB pathway. Thus,. the current study provided some clarity in terms of understanding the immune response about rrad gene differences between disease-resistant and disease-susceptible individuals. This study provides a molecular basis for fish gene functional analysis and may serve as a reference for in-depth of bacterial disease resistance of teleost.
与糖尿病相关的 Ras（RRAD）是 Ras GTPase 超家族的成员，在许多细胞功能中发挥作用，如细胞增殖和分化。特别是，该超家族作为 NF-κB 信号通路抑制剂和钙调节剂参与免疫反应途径。最近的转录组研究表明，与易感个体相比，抗病性日本牙鲆个体的脾脏中表达了 ，qPCR 结果也验证了这一点。因此，本研究旨在探讨 如何通过 NF-κB 途径调节抗菌免疫。首先，鉴定了 的编码序列。该序列长 1092bp，编码 364 个氨基酸。基于 和结构关系分析， 似乎与硬骨鱼关系更密切。接下来，评估了免疫相关组织中抗病性和易感性个体之间的 表达差异，结果表明 主要在抗病性个体的脾脏中表达。在 感染后，脾脏中的 表达发生变化。在体外，共培养评估了抗病性脾脏中 启动子的低甲基化水平，这与高 mRNA 表达一致。用 gill 细胞系对 进行 siRNA 介导的敲低，影响了许多 -network 相关基因，如 、 、 、 、 、 、 、 、 、 和其他一些炎症相关基因，如 。此外，流式细胞术分析表明， 过表达更可能诱导细胞凋亡，建立了 与调节 NF-κB 途径的功能之间的联系。因此，RRAD 基因在抗病性和易感性 个体之间的免疫反应差异方面提供了一些清晰度。本研究为鱼类 基因功能分析提供了分子基础，并可为深入研究硬骨鱼的细菌抗病性提供参考。
