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GFP 转基因斑马鱼(Oryzias latipes)在诱导型 cyp1a 启动子的作用下,为 TCDD 和其他持久性有机污染物的存在提供了一种敏感、便捷的生物指示剂。

GFP transgenic medaka (Oryzias latipes) under the inducible cyp1a promoter provide a sensitive and convenient biological indicator for the presence of TCDD and other persistent organic chemicals.

机构信息

Department of Biological Sciences, NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore.

出版信息

PLoS One. 2013 May 20;8(5):e64334. doi: 10.1371/journal.pone.0064334. Print 2013.

DOI:10.1371/journal.pone.0064334
PMID:23700472
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3659123/
Abstract

Persistent organic pollutants (POPs) are resistant to environmental degradation and can cause multitude of health problems. Cytochrome P450 1A (Cyp1a) is often up-regulated by POPs through the activation of aryl hydrocarbon receptor (AhR) pathway and is thus usually used as a biomarker for xenobiotics exposure. To develop a convenient in vivo tool to monitor xenobiotic contamination in the water, we have established GFP transgenic medaka using the inducible cyp1a promoter, Tg(cyp1a:gfp). Here we tested Tg(cyp1a:gfp) medaka at three different stages, prehatching embryos, newly hatched fry and adult with 2,3,7,8-tetrachlorodiebnzo-p-dioxin (TCDD), a dioxin. While GFP induction was observed in all three stages, newly hatched fry were the most sensitive with the lowest observed effective concentration of 0.005 nM or 16.1 ng/L. The highly sensitive organs included the kidney, liver and intestine. With high concentrations of TCDD, several other organs such as the olfactory pit, tail fin, gills, lateral line neuromast cells and blood vessels also showed GFP expression. In addition, Tg(cyp1a:gfp) medaka fry also responded to two other AhR agonists, 3-methylcholanthrene and benzo[a]pyrene, for GFP induction, but no significant GFP induction was observed towards several other chemicals tested, indicating the specificity of this transgenic line. The GFP inducibility of Tg(cyp1a:gfp) medaka at both fry and adult stages may be useful for development of high-throughput assays as well as online water monitoring system to detect xenobiotic toxicity.

摘要

持久性有机污染物 (POPs) 具有抗环境降解性,并可能导致多种健康问题。细胞色素 P450 1A(Cyp1a)通常通过芳香烃受体(AhR)途径的激活而上调,因此通常用作外源性物质暴露的生物标志物。为了开发一种方便的体内工具来监测水中的外源性污染物,我们使用可诱导的 cyp1a 启动子 Tg(cyp1a:gfp) 建立了 GFP 转基因斑马鱼。在这里,我们在三个不同阶段测试了 Tg(cyp1a:gfp) 斑马鱼,即孵化前的胚胎、刚孵化的幼鱼和成年鱼,使用的是 2,3,7,8-四氯二苯并对二恶英(TCDD),一种二恶英。虽然在所有三个阶段都观察到 GFP 诱导,但刚孵化的幼鱼最敏感,最低观察到的有效浓度为 0.005 nM 或 16.1ng/L。高度敏感的器官包括肾脏、肝脏和肠道。在高浓度的 TCDD 下,嗅觉坑、尾鳍、鳃、侧线神经嵴细胞和血管等其他几个器官也显示出 GFP 表达。此外,Tg(cyp1a:gfp) 斑马鱼幼鱼对另外两种 AhR 激动剂 3-甲基胆蒽和苯并[a]芘也有 GFP 诱导反应,但对测试的其他几种化学物质没有明显的 GFP 诱导反应,表明该转基因系具有特异性。Tg(cyp1a:gfp) 斑马鱼幼鱼和成年鱼阶段的 GFP 可诱导性可能有助于开发高通量测定法以及在线水监测系统,以检测外源性毒性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/6eb3d6fdad2b/pone.0064334.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/24b82abe7b7c/pone.0064334.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/200e7c2f2c7f/pone.0064334.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/70157aca000b/pone.0064334.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/fc7a3e8453f3/pone.0064334.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/6eb3d6fdad2b/pone.0064334.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/24b82abe7b7c/pone.0064334.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/200e7c2f2c7f/pone.0064334.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/70157aca000b/pone.0064334.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/fc7a3e8453f3/pone.0064334.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/3659123/6eb3d6fdad2b/pone.0064334.g005.jpg

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