Aquatic Toxicology：臺灣師大 | AgNPs和CuNPs抑制斑馬魚胚胎側線毛細胞鈣吸收（附NMT實驗體系）

NMT是基因功能的活體檢測技術，已被103位諾貝爾獎得主所在單位，及北大、清華、中科院使用。

期刊：Aquatic Toxicology
主題：AgNPs和CuNPs抑制斑馬魚胚胎側線毛細胞鈣吸收

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

影響因子：3.794
檢測指標：Ca2+流速

檢測樣品：斑馬魚胚胎側線毛細胞發束

Ca2+流實驗處理方法：

斑馬魚胚胎經0-96hpf受精后，分別用0，0.1，1，3ppm的AgNPs和0，0.01，0.1，1ppm的CuNPs進行受精
Ca2+流實驗測試液成份：
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：臺灣師范大學林豊益

斑馬魚毛細胞離子流檢測

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（谷歌機翻）

納米粒子（NPs）對魚類早期的潛在毒性仍不清楚。在這項研究中，我們調查了銀（AgNPs）和銅納米顆粒（CuNPs）對斑馬魚胚胎側線毛細胞的毒性作用。

斑馬魚胚胎在受精后0?96 h（hpf）的不同濃度的AgNPs和CuNPs中孵育。發現AgNPs和CuNPs均以劑量依賴性方式引起斑馬魚胚胎的毒性作用。AgNP和CuNP的96小時50％致死濃度（LC50）值分別為6.1 ppm（56.5μM）和2.61 ppm（41.1μM）。AgNPs [≥1ppm（9.3μM）]和CuNPs [≥0.01ppm（0.16μM）]嚴重損害了FM1-43標記的毛細胞數量和發束的微觀結構。

用掃描離子選擇微電極技術測量在毛細胞的發束處的Ca 2+流入，以評估毛細胞的功能。發現AgNPs [≥0.1ppm（0.9μM）]和CuNPs [≥0.01ppm（0.16μM）]均能顯著減少Ca2 +的流入。在暴露于AgNPs和CuNPs 4小時（96?100 hpf）的孵化胚胎中也發現了類似的毒性作用。

這項研究表明，斑馬魚的側線毛細胞對AgNPs和CuNPs敏感，而這些污染物在水生環境中可能對魚類的生存構成威脅。

結果表明：在AgNP處理組中，相比于0 ppm，Ca2+吸收速率在0.1,1和3 ppm時分別減少了26%,46%和91%;在CuNP處理組中，相比于0 ppm，Ca2+吸收速率在0.01 ppm時減少了38%，而在0.1和1 ppm時幾乎檢測不到Ca2+的吸收速率。由此可見，AgNPs （≥0.1 ppm）和 CuNPs（≥0.01ppm）都會顯著抑制Ca2+的吸收。海洋環境中的污染物會威脅魚類的生存。