Molecular characterization of ten zinc (Zn) transporter genes and their regulation to Zn metabolism in freshwater teleost yellow catfish Pelteobagrus fulvidraco
Molecular characterization of ten zinc (Zn) transporter genes and their regulation to Zn metabolism in freshwater teleost yellow catfish Pelteobagrus fulvidraco
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淡水硬骨鱼黄颡鱼十个锌（Zn）转运蛋白基因的分子特征及其对锌代谢的调控
DOI：
10.1016/j.jtemb.2019.126433
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发表时间：
2020-05-01
影响因子：
3.5
通讯作者：
Luo, Zhi
中科院分区：
文献类型：
--
作者：
Chen, Shu-Wei;Wu, Kun;Luo, Zhi
背景：锌是脊椎动物必需的微量元素，锌的摄取和转运与锌转运蛋白ZIP家族有关。方法：从黄颡鱼（Pelteobagrus fulvidraco）中克隆了10个锌转运相关基因（ZIP 1、ZIP 3、ZIP 6、ZIP 7、ZIP 8、ZIP 9、ZIP 10、ZIP 11、ZIP 13和ZIP 14）的cDNA全长序列，并对其mRNA组织表达进行了研究。这些ZIP mRNA的表达也进行了评估，在原代肝细胞和肠上皮细胞的黄颡鱼响应三个锌水平（0，30 μ M和60 μ M，分别）。10个ZIP基因的mRNA表达在9个供试组织中检测到，但这些组织之间的变化。流式细胞仪分析和共聚焦显微镜观察结果表明，在24 h和48 h，肝细胞和肠上皮细胞内游离Zn 2+浓度均随Zn孵育浓度的增加而增加。锌孵育差异影响ZIP转运蛋白在肝细胞和肠上皮细胞的mRNA水平，在时间和细胞依赖性的方式。在肝细胞中，24 h时，与对照组相比，Zn添加下调ZIP 1，ZIP 3，ZIP 6，ZIP 7，ZIP 8，ZIP 9，ZIP 11和ZIP 14的mRNA水平;然而，ZIP 10 mRNA水平在60 μ M Zn组低于对照组和30 μ M Zn组。在48 h时，ZIP 1、ZIP 6、ZIP 7、ZIP 9、ZIP 10和ZIP 14的mRNA水平随Zn孵育浓度的增加而下降; ZIP 3的mRNA水平在60 μ M Zn组最低，与对照组和30 μ M Zn组无显著差异。在肠上皮细胞中，24 h时，Zn处理组ZIP 1、ZIP 6、ZIP 7、ZIP 8、ZIP 9、ZIP 10、ZIP 11、ZIP 13和ZIP 14的mRNA水平均显著下降，60 μ M Zn处理组ZIP 3的mRNA水平低于对照组和30 μ M Zn处理组。在48 h时，Zn处理上调了ZIP 6和ZIP 9的mRNA水平，下调了ZIP 8、ZIP 10和ZIP 13的mRNA水平。ZIP 7、ZIP 11和ZIP 14 mRNA丰度在60 μ M Zn组中最低，与对照组和30 μ M Zn组相比差异不显著。结论：本研究首次鉴定了黄颡鱼ZIP家族10个成员，并探讨了它们的mRNA组织表达。在黄颡鱼的肝细胞和肠上皮细胞中研究了它们对锌添加的调节作用。我们的研究揭示了细胞暴露于锌的机制，并提供了新的见解锌稳态的调节机制。
Background: Zn is an essential trace element for vertebrates, and Zn uptake and transport is related with the ZIP family of Zn transporters. Meantime, Zn also influenced the expression of ZIP family members.Methods: We cloned and characterized the full-length cDNA sequences of ten Zn transport-relevant genes (ZIP1, ZIP3, ZIP6, ZIP7, ZIP8, ZIP9, ZIP10, ZIP11, ZIP13 and ZIP14) from yellow catfish Pelteobagrus fulvidraco, investigated their mRNA tissue expression. These ZIP mRNA expression was also assessed in the primary hepatocytes and intestinal epithelial cells of yellow catfish in response to three Zn levels (0, 30 mu M and 60 mu M, respectively).Results: All these genes shared the similar domains with the corresponding members in mammals. The mRNA expression of the ten ZIP genes was detected in nine-tested tissues, but variable among these tissues. Flow cytometry analysis and confocal microscopy observation indicated that intracellular free Zn2+ concentration in hepatocytes and intestinal epithelial cells increased with increasing Zn incubation concentration at both 24 h and 48 h. Zn incubation differentially influenced mRNA levels of ZIP transporters in the hepatocytes and intestinal epithelial cells, in a time- and cells-dependent manners. In the hepatocytes, at 24 h, compared to the control, Zn addition down-regulated mRNA levels of ZIP1, ZIP3, ZIP6, ZIP7, ZIP8, ZIP9, ZIP11 and ZIP14; however, ZIP10 mRNA levels were lower in 60 mu M Zn group than those in the control and 30 mu M Zn group. At 48 h, mRNA levels of ZIP1, ZIP6, ZIP7, ZIP9, ZIP10 and ZIP14 declined with increasing Zn incubation concentrations; ZIP3 mRNA levels were the lowest in 60 mu M Zn group and showed no significant differences between the control and 30 mu M Zn group. In the intestinal epithelial cells, at 24 h, Zn addition down-regulated mRNA levels of ZIP1, ZIP6, ZIP7, ZIP8, ZIP9, ZIP10, ZIP11, ZIP13 and ZIP14; ZIP3 mRNA levels were lower in 60 mu M Zn group than those in the control and 30 mu M Zn group. At 48 h, Zn addition up-regulated mRNA levels of ZIP6 and ZIP9, but down-regulated mRNA levels of ZIP8, ZIP10 and ZIP13. ZIP7, ZIP11 and ZIP14 mRNA abundances were the lowest in 60 mu M Zn group and showed no significant differences between the control and 30 mu M Zn group.Conclusion: For the first time, our study characterized ten ZIP family members in yellow catfish, explored their mRNA tissue expression. Their regulation to Zn addition were also investigated in the hepatocytes and intestinal epithelial cells of yellow catfish. Our study revealed the mechanism of cells exposed to Zn addition and provided novel insights for the regulatory mechanism of Zn homeostasis.