Aquatic Toxicology：台湾师大 | AgNPs和CuNPs抑制斑马鱼胚胎侧线毛细胞钙吸收（附NMT实验体系）

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。

期刊：Aquatic Toxicology
主题：AgNPs和CuNPs抑制斑马鱼胚胎侧线毛细胞钙吸收

标题：Toxic e?ects of silver and copper nanoparticles on lateral-line hair cells of zebra?sh embryos

影响因子：3.794
检测指标：Ca2+流速

检测样品：斑马鱼胚胎侧线毛细胞发束

Ca2+流实验处理方法：

斑马鱼胚胎经0-96hpf受精后，分别用0，0.1，1，3ppm的AgNPs和0，0.01，0.1，1ppm的CuNPs进行受精
Ca2+流实验测试液成份：
0.5mM NaCl，0.2mM CaSO4，0.2mM MgSO4，0.16mM KH2PO4，and0.16mM K2HPO4（pH 7.0），300μM MOPS，0.1mg/l tricaine

作者：台湾师范大学林豊益

斑马鱼毛细胞离子流检测

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 dosedependent 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 lateralline 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仍不清楚。在这项研究中，我们调查了银（AgNPs）和铜纳米颗粒（CuNPs）对斑马鱼胚胎侧线毛细胞的毒性作用。

斑马鱼胚胎在受精后0?96   h（hpf）的不同浓度的AgNPs和CuNPs中孵育。发现AgNPs和CuNPs均以剂量依赖性方式引起斑马鱼胚胎的毒性作用。AgNP和CuNP的96小时50％致死浓度（LC50）值分别为6.1  ppm（56.5μM）和2.61 ppm（41.1μM）。AgNPs [≥1ppm（9.3μM）]和CuNPs   [≥0.01ppm（0.16μM）]严重损害了FM1-43标记的毛细胞数量和发束的微观结构。

用扫描离子选择微电极技术测量在毛细胞的发束处的Ca  2+流入，以评估毛细胞的功能。发现AgNPs [≥0.1ppm（0.9μM）]和CuNPs  [≥0.01ppm（0.16μM）]均能显着减少Ca2 +的流入。在暴露于AgNPs和CuNPs 4小时（96?100   hpf）的孵化胚胎中也发现了类似的毒性作用。

这项研究表明，斑马鱼的侧线毛细胞对AgNPs和CuNPs敏感，而这些污染物在水生环境中可能对鱼类的生存构成威胁。

结果表明：在AgNP处理组中，相比于0 ppm，Ca2+吸收速率在0.1,1和3   ppm时分别减少了26%,46%和91%;在CuNP处理组中，相比于0 ppm，Ca2+吸收速率在0.01 ppm时减少了38%，而在0.1和1  ppm时几乎检测不到Ca2+的吸收速率。由此可见，AgNPs （≥0.1  ppm）和 CuNPs（≥0.01ppm）都会显著抑制Ca2+的吸收。海洋环境中的污染物会威胁鱼类的生存。