GATA-3 in Atlantic salmon (Salmo salar): Tissue distribution and its regulation of IL-4/13a promoter.
GATA-3 in Atlantic salmon (Salmo salar): Tissue distribution and its regulation of IL-4/13a promoter.
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大西洋鲑鱼 (Salmo salar) 中的 GATA-3：组织分布及其对 IL-4/13a 启动子的调控
DOI：
10.3389/fcimb.2022.1063600
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发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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作者：
GATA 3是一种转录因子，在T细胞谱系分化和辅助性T细胞2（Th 2）型免疫应答中起重要作用。在这项研究中，我们开发了两种抗大西洋鲑鱼加塔-3的大鼠抗体（分别为抗rSs加塔-3a和抗rSs加塔-3b）。免疫印迹和免疫荧光结果表明，抗rSsGATA-3b抗体可识别内源性SsGATA-3蛋白，而抗rSsGATA-3a抗体不与SsGATA-3结合。免疫组织化学分析显示，SsGATA-3阳性细胞检测到所有组织中测试，与相对较高数量的免疫反应细胞在鳃和脾脏。免疫组化结果显示，SsGATA-3在柱细胞、上皮细胞、软骨细胞、软骨膜细胞和一些未分化的基底细胞中均有表达。此外，我们确定了577 bp的上游启动子序列的SsIL-4/13 a，并发现四个基序匹配的SsGATA-3结合位点。通过检测4个缺失报告基因构建体和SsGATA-3过表达质粒，对SsIL-4/13 a启动子区进行了分析。结果表明，SsGATA-3在转录起始位点上游-317 ~-302 bp区域内增强了SsIL-4/13 a启动子的活性。针对Th 2标志物如加塔-3的抗体在解决鱼类中T细胞应答的多样性方面是有价值的。
GATA3 is a transcription factor that plays an important role in T cell lineage differentiation and T-helper 2 (Th2) type immune responses. In this study, we developed two rat antibodies against Atlantic salmon GATA-3 (anti-rSsGATA-3a and anti-rSsGATA-3b, respectively). The western blotting and immunofluorescence results showed that anti-rSsGATA-3b antibodies recognized endogenous SsGATA-3 proteins, while the anti-rSsGATA-3a antibodies did not bind SsGATA-3. Immunohistochemical analysis revealed that SsGATA-3 positive cells were detected in all tissues tested, with relatively high number of immune reactive cells in the gills and spleen. Furthermore, the immunohistochemical study revealed that SsGATA-3 was expressed in pillar cells, epithelial cells, chondrocytes, perichondrium cells, and some undifferentiated basal cells. In addition, we determined 577 bp of the upstream promoter sequence of SsIL-4/13a and found four motifs that matched SsGATA-3 binding sites. The promoter regions of SsIL-4/13a were assessed by transfecting four deletion reporter constructs and SsGATA-3 overexpression plasmids. The result showed that SsGATA-3 enhanced the activity of SsIL-4/13a promoters within the region ranging from -317 to -302 bp upstream of the transcriptional start site. Antibodies against Th2 markers such as GATA-3 are valuable in addressing the diversity of T cell responses in fish.
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