期刊：Aquatic Toxicology

主题：银铜纳米颗粒对斑马鱼胚胎侧线毛细胞的毒性作用

标题：Toxic effects of silver and copper nanoparticles on lateral-line hair cells of
zebrafish embryos

影响因子：3.794

检测指标：Ca2+流速

检测部位：斑马鱼胚胎侧线毛细胞

Ca2+流速流实验处理方法：

斑马鱼胚胎在受精后0-96 h（hpf），0, 0.9, 9.3, 27.8 μM的AgNPs，和0,0.16,1.6，15.8μM的CuNP分别进行处理

Ca2+流速流实验测试液成份：

0.5mM NaCl, 0.2mM CaSO4, 0.2mM MgSO4, 0.16mM KH2PO4, and 0.16mM K2HPO4 (pH 7.0)，300μM MOPS，0.1 mg/l tricaine

作者：台湾师范大学林豊益、Hsiu-Ju Yen

英文摘要

The potential toxicity of nanoparticles (NPs) to the early stages of fish is still unclear. In this study, we investigated the toxic effects of silver (AgNPs) and copper nanoparticles (CuNPs) on lateral-line hair cells of zebrafish embryos.

Zebrafish embryos were incubated in different concentrations of AgNPs and CuNPs at 0˜96 h post-fertilization (hpf). Both AgNPs and CuNPs were found to cause toxic effects in zebrafish embryos in a dose-dependent manner. Values of the 96-h 50% lethal concentration (LC50) of AgNPs and CuNPs were 6.1 ppm (56.5 μM) and 2.61 ppm (41.1 μM), respectively.

The number of FM1-43-labeled hair cells and the microstructure of hair bundles were significantly impaired by AgNPs [≥1 ppm (9.3 μM)] and CuNPs [≥0.01 ppm (0.16 μM)]. Ca2+ influxes at hair bundles of hair cells were measured with a scanning ion-selective microelectrode technique to evaluate the function of hair cells.

AgNPs [≥0.1 ppm (0.9 μM)] and CuNPs [≥0.01 ppm (0.16 μM)] were both found to significantly reduce Ca2+ influxes. Similar toxic effects were also found in hatched embryos subjected to 4 h of exposure (96˜100 hpf) to AgNPs and CuNPs. This study revealed that lateral-line hair cells of zebrafish are susceptible to AgNPs and CuNPs, and these contaminants in aquatic environments could pose a threat to fish survival.

中文摘要（谷歌机翻）

纳米粒子（NPs）对鱼类早期的潜在毒性尚不清楚。在这项研究中，我们研究了银（AgNPs）和铜纳米颗粒（CuNPs）对斑马鱼胚胎侧线毛细胞的毒性作用。

在受精后0~96h（hpf），将斑马鱼胚胎在不同浓度的AgNPs和CuNPs中孵育。发现AgNPs和CuNPs都以剂量依赖的方式对斑马鱼胚胎产生毒性作用。 AgNPs和CuNPs的96小时50％致死浓度（LC50）的值分别为6.1ppm（56.5μM）和2.61ppm（41.1μM）。

AgNPs [≥1ppm（9.3μM）]和CuNPs [≥0.01ppm（0.16μM）]显着损害了FM1-43标记的毛细胞数量和发束的微观结构。用扫描离子选择性微电极技术测量毛细胞发束处的Ca2+流入，以评估毛细胞的功能。

发现AgNPs [≥0.1ppm（0.9μM）]和CuNPs [≥0.01ppm（0.16μM）]均可显着降低Ca2+流入量。在孵化4h（96〜100hpf）的AgNPs和CuNPs孵化的胚胎中也发现了类似的毒性效应。这项研究表明，斑马鱼的侧线毛细胞对AgNPs和CuNPs敏感，这些污染物在水生环境中可能对鱼类的存活构成威胁。

结果表明：在经历96小时AgNP或CuNP处理的胚胎中也发现了毛细胞的功能损伤。使用NM测量L1神经瘤的毛细胞的Ca2+流入量（图A），在0.1,1和3ppm（0.9,9.3和27.8μM）的AgNP组中，Ca2+流入量显着减少26％，46％和91％（图B）。在0.01 ppm（0.16μM）CuNP组中，Ca2+流入量减少了38％，而在0.1和1 ppm（1.6和15.8μM）CuNP组中，Ca2+流入几乎检测不到（图C）

联盟专家暨本文作者介绍

林豊益，博士，教授。台湾师范大学生命科学学院生理组，主要从事鱼类环境生理、水生环境毒理、机械性感觉生理、斑马鱼疾病模式与药物筛选研究。

近期代表性成果

• Lin, L. Y., Y. H. Yeh, G.Y. Hung, C. H. Lin, P. P. Hwang, J. L. Horng. 2018. Role of calcium-sensingreceptor in mechanotransducer-channel-mediated Ca2+ influx in hair cells ofzebrafish larvae. Front. Physiol. 649:9.
• L. Horng, L. L. Yu, S. T. Liu, P. Y. Chen, L. Y. Lin. 2017. Potassiumregulation in medaka (Oryzias latipes) larvae acclimated to fresh water:passive uptake and active secretion by the skin cells. Sci. Rep. 7, 16215.
• Liu, S. T., J. L. Horng, P. Y. Chen, P. P. Hwang, L.Y. Lin. 2016. Saltsecretion is linked to acid-base regulation of ionocytes in seawater-acclimatedmedaka: new insights into the salt-secreting mechanism. Sci. Rep. 6, 31433.
• Horng, J. L., P. L. Chao, P. Y. Chen, T. H. Shih, L.Y. Lin. 2015.Aquaporin 1 is involved in acid secretion by ionocytes of zebrafish embryosthrough facilitating CO2 transport. Plos One 10(8):e0136440.
• Lin, Y. H., G. Y. Hung, L. C. Wu, S. W. Chen, L. Y. Lin. J. L. Horng.2015. Anion exchanger 1b in sereocilia is required for the functioning ofmechanotransducer channels in lateral-line hair cells of zebrafish. Plos One10(2): e0117041.
  
  
  洪君琳，博士，副教授。台北医学大学医学院医学科学研究所，斑马鱼模式核心服务实验室主任，主要从事以斑马鱼为模式研究毛细胞功能及药物影响的研究。

  近期代表性成果

• Lin LY, et al,. Role of Calcium-Sensing Receptor in Mechanotransducer Channel Mediated Ca2+ Influx in Hair Cells of Zebrafish Larvae . frontiers in physiology .2018 ;(9):649

• Hung Giun Yi, et al,. Incidence of immune thrombocytopenia in Taiwan: a nationwide population-based study . Transfusion .2018 ;(58):2712-2719

• Horng JL, et al,. Potassium Regulation in Medaka (Oryzias latipes) Larvae Acclimated to Fresh Water: Passive Uptake and Active Secretion by the Skin Cells. . Sci Rep. .2017 ;(7):16215

• Hung GY, et al,. Cancer in adolescents: Incidences and trends during 1995-2009 in Taiwan. . Cancer Lett .2016 ;(372):110-117

• Hung GY, et al,. Changing incidence patterns of hepatocellular carcinoma among age groups in Taiwan . J Hepatol .2015 ;(63):1390-1396

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