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期刊：Aquatic Toxicology

主題：銀銅納米顆粒對斑馬魚胚胎側(cè)線毛細(xì)胞的毒性作用

標(biāo)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(biāo)：Ca2+流速

檢測部位：斑馬魚胚胎側(cè)線毛細(xì)胞

Ca2+流速流實驗處理方法：

斑馬魚胚胎在受精后0-96 h（hpf），0, 0.9, 9.3, 27.8 μM的AgNPs，和0,0.16,1.6，15.8μM的CuNP分別進(jìn)行處理

Ca2+流速流實驗測試液成份：

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：中國臺灣師范大學(xué)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（谷歌機翻）

納米粒子（NPs）對魚類早期的潛在毒性尚不清楚。在這項研究中，我們研究了銀（AgNPs）和銅納米顆粒（CuNPs）對斑馬魚胚胎側(cè)線毛細(xì)胞的毒性作用。

 在受精后0~96h（hpf），將斑馬魚胚胎在不同濃度的AgNPs和CuNPs中孵育。發(fā)現(xiàn)AgNPs和CuNPs都以劑量依賴的方式對斑馬魚胚胎產(chǎn)生毒性作用。 AgNPs和CuNPs的96小時50％致死濃度（LC50）的值分別為6.1ppm（56.5μM）和2.61ppm（41.1μM）。

AgNPs [≥1ppm（9.3μM）]和CuNPs [≥0.01ppm（0.16μM）]顯著損害了FM1-43標(biāo)記的毛細(xì)胞數(shù)量和發(fā)束的微觀結(jié)構(gòu)。用掃描離子選擇性微電極技術(shù)測量毛細(xì)胞發(fā)束處的Ca2+流入，以評估毛細(xì)胞的功能。

發(fā)現(xiàn)AgNPs [≥0.1ppm（0.9μM）]和CuNPs [≥0.01ppm（0.16μM）]均可顯著降低Ca2+流入量。在孵化4h（96?100hpf）的AgNPs和CuNPs孵化的胚胎中也發(fā)現(xiàn)了類似的毒性效應(yīng)。這項研究表明，斑馬魚的側(cè)線毛細(xì)胞對AgNPs和CuNPs敏感，這些污染物在水生環(huán)境中可能對魚類的存活構(gòu)成威脅。

結(jié)果表明：在經(jīng)歷96小時AgNP或CuNP處理的胚胎中也發(fā)現(xiàn)了毛細(xì)胞的功能損傷。使用NM測量L1神經(jīng)瘤的毛細(xì)胞的Ca2+流入量（圖A），在0.1,1和3ppm（0.9,9.3和27.8μM）的AgNP組中，Ca2+流入量顯著減少26％，46％和91％（圖B）。在0.01 ppm（0.16μM）CuNP組中，Ca2+流入量減少了38％，而在0.1和1 ppm（1.6和15.8μM）CuNP組中，Ca2+流入幾乎檢測不到（圖C）

聯(lián)盟專家暨本文作者介紹

林豊益，博士，教授。中國臺灣師范大學(xué)生命科學(xué)學(xué)院生理組，主要從事魚類環(huán)境生理、水生環(huán)境毒理、機械性感覺生理、斑馬魚疾病模式與藥物篩選研究。

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。臺北醫(yī)學(xué)大學(xué)醫(yī)學(xué)院醫(yī)學(xué)科學(xué)研究所，斑馬魚模式核心服務(wù)實驗室主任，主要從事以斑馬魚為模式研究毛細(xì)胞功能及藥物影響的研究。

• 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