Exposure to tributyltin induces endoplasmic reticulum stress and the unfolded protein response in zebrafish
Exposure to tributyltin induces endoplasmic reticulum stress and the unfolded protein response in zebrafish
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DOI：
10.1016/j.aquatox.2013.08.017
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发表时间：
2013-10-15
影响因子：
4.5
通讯作者：
Matsuoka, Masato
中科院分区：
文献类型：
--
作者：
Komoike, Yuta;Matsuoka, Masato
三丁基锡（TBT）是一种主要的海洋污染物，可引起内分泌干扰、肝毒性、免疫毒性和神经毒性。然而，三丁基锡化合物毒性的分子机制尚未完全阐明。我们研究是否暴露于TBT诱导内质网（ER）应激反应的模式生物斑马鱼。斑马鱼来源的BRF 41成纤维细胞暴露于0.5或1 μ M TBT 0.5-16小时，随后裂解和免疫印迹检测ER应激相关蛋白。斑马鱼胚胎，直到32小时后受精（hpf），暴露于1 μ M TBT为16小时，并在整个安装在原位杂交和免疫组化可视化ER伴侣和ER应力相关的凋亡因子的表达。BRF 41细胞暴露于三丁基锡化合物后，引起斑马鱼蛋白激酶RNA激活样雌激素受体激酶（PERK）、真核生物翻译起始因子2 α（eIF 2 α）和肌醇需要酶1（IRE 1）的同源物磷酸化，X盒结合蛋白1（XBP 1）mRNA的特征性剪接，以及转录激活因子4（ATF 4）蛋白的表达增强。在TBT暴露的斑马鱼胚胎，异位表达的基因编码斑马鱼同源的78 kDa的葡萄糖调节蛋白（GRP 78）和基因编码CCAAT/增强子结合蛋白同源蛋白（CHOP）的前体中检测到的神经肥大，这是一个感觉器官，用于检测水流和振动。我们的体外和体内研究表明，斑马鱼暴露于TBT诱导ER应激反应，通过激活PERK-eIF 2 α和IRE 1-XBP 1途径的未折叠蛋白反应（UPR）在器官特异性的方式。(C)2013爱思唯尔有限公司版权所有。
Tributyltin (TBT) is a major marine contaminant and causes endocrine disruption, hepatotoxicity, immunotoxicity, and neurotoxicity. However, the molecular mechanisms underlying the toxicity of TBT have not been fully elucidated. We examined whether exposure to TBT induces the endoplasmic reticulum (ER) stress response in zebrafish, a model organism. Zebrafish-derived BRF41 fibroblast cells were exposed to 0.5 or 1 mu M TBT for 0.5-16 h and subsequently lysed and immunoblotted to detect ER stress-related proteins. Zebrafish embryos, grown until 32 h post fertilization (hpf), were exposed to 1 mu M TBT for 16 h and used in whole mount in situ hybridization and immunohistochemistry to visualize the expression of ER chaperones and an ER stress-related apoptosis factor. Exposure of the BRF41 cells to TBT caused phosphorylation of the zebrafish homolog of protein kinase RNA-activated-like ER kinase (PERK), eukaryotic translation initiation factor 2 alpha (eIF2 alpha), and inositol-requiring enzyme 1 (IRE1), characteristic splicing of X-box binding protein 1 (XBP1) mRNA, and enhanced expression of activating transcription factor 4 (ATF4) protein. In TBT-exposed zebrafish embryos, ectopic expression of the gene encoding zebrafish homolog of the 78 kDa glucose-regulating protein (GRP78) and gene encoding CCAAT/enhancer-binding protein homologous protein (CHOP) was detected in the precursors of the neuromast, which is a sensory organ for detecting water flow and vibration. Our in vitro and in vivo studies revealed that exposure of zebrafish to TBT induces the ER stress response via activation of both the PERK-eIF2 alpha and IRE1-XBP1 pathways of the unfolded protein response (UPR) in an organ-specific manner. (C) 2013 Elsevier B.V. All rights reserved.