Using Electroporation to Improve and Accelerate Zebrafish Embryo Toxicity Testing.
Using Electroporation to Improve and Accelerate Zebrafish Embryo Toxicity Testing.
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DOI：
10.3390/mi15010049
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发表时间：
2023-12-26
期刊：
影响因子：
3.4
通讯作者：
中科院分区：
文献类型：
--
作者：
斑马鱼已成为生物医学研究的有用模型，并已用于环境毒理学研究。然而，绒毛膜在胚胎阶段的存在限制了细胞对有毒元素的暴露，并在鱼胚胎毒性测试（FET）中产生假阴性或降低敏感性的可能性。本文介绍了使用电穿孔作为一种技术来改善绒毛膜内有毒元素的输送，增加毒素在胚胎早期（受精后<3小时）的暴露水平。一种定制的带有所需电路的电穿孔装置已经开发出来，可以将胚胎放置在提供电脉冲以加速分子进入绒毛膜的电极之间。优化后的参数有利于材料进入绒毛膜，而不影响胚胎的存活率。利用台锥蓝染料和金纳米颗粒（AuNPs, 20-40 nm）证明了电穿孔系统的有效性。我们的研究结果证明了通过优化电学参数（包括脉冲宽度、脉冲数和振幅）来控制绒毛膜内传递的染料和纳米颗粒浓度的可行性。接下来，我们测试了银纳米颗粒（AgNPs， 10纳米），这是一种常用的毒素，可以降低死亡率，影响心率，并引起表型缺陷。我们发现AgNPs的电穿孔将毒性测试所需的暴露时间从4天减少到几个小时。FET的电穿孔可以使潜在毒素快速进入斑马鱼胚胎，减少毒性测试和给药实验所需的时间。
Zebrafish have emerged as a useful model for biomedical research and have been used in environmental toxicology studies. However, the presence of the chorion during the embryo stage limits cellular exposure to toxic elements and creates the possibility of a false-negative or reduced sensitivity in fish embryo toxicity testing (FET). This paper presents the use of electroporation as a technique to improve the delivery of toxic elements inside the chorion, increasing the exposure level of the toxins at an early embryo stage (<3 h post-fertilization). A custom-made electroporation device with the required electrical circuitry has been developed to position embryos between electrodes that provide electrical pulses to expedite the entry of molecules inside the chorion. The optimized parameters facilitate material entering into the chorion without affecting the survival rate of the embryos. The effectiveness of the electroporation system is demonstrated using Trypan blue dye and gold nanoparticles (AuNPs, 20–40 nm). Our results demonstrate the feasibility of controlling the concentration of dye and nanoparticles delivered inside the chorion by optimizing the electrical parameters, including pulse width, pulse number, and amplitude. Next, we tested silver nanoparticles (AgNPs, 10 nm), a commonly used toxin that can lower mortality, affect heart rate, and cause phenotypic defects. We found that electroporation of AgNPs reduces the exposure time required for toxicity testing from 4 days to hours. Electroporation for FET can provide rapid entry of potential toxins into zebrafish embryos, reducing the time required for toxicity testing and drug delivery experiments.
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通过电穿孔在成年斑马鱼中转基因的有效表达。
影响因子：
3.5
作者：
Rambabu, KM;Rao, SHN;Rao, NM
通讯作者：
Rao, NM
DOI：
10.1016/j.devbrainres.2004.03.016
发表时间：
2004-07-19
期刊：
DEVELOPMENTAL BRAIN RESEARCH
影响因子：
--
作者：
Creton, R
通讯作者：
Creton, R
影响因子：
4.1
作者：
Cheng, Jinping;Flahaut, Emmanuel;Cheng, Shuk Han
通讯作者：
Cheng, Shuk Han
影响因子：
6.1
作者：
Choudhury, Deepak;van Noort, Danny;Yu, Hanry
通讯作者：
Yu, Hanry
影响因子：
5
作者：
Asharani, P. V.;Yi Lianwu;Valiyaveettil, Suresh
通讯作者：
Valiyaveettil, Suresh