Application of transgenic zebrafish for investigating inflammatory responses to nanomaterials: Recommendations for new users
Application of transgenic zebrafish for investigating inflammatory responses to nanomaterials: Recommendations for new users
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DOI：
10.12688/f1000research.128851.1
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发表时间：
2023-01
期刊：
影响因子：
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通讯作者：
Helinor J Johnston;S. Gillies;R. Verdon;V. Stone;T. Henry;L. Tran;C. Tucker;A. Rossi;C. Tyler
中科院分区：
文献类型：
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作者：
Helinor J Johnston;S. Gillies;R. Verdon;V. Stone;T. Henry;L. Tran;C. Tucker;A. Rossi;C. Tyler
尽管在一系列消费品中越来越多地使用纳米材料，但其对人类健康的潜在不利影响仍存在不确定性。NM是否激活促炎反应的研究通常用于评估其在体外和体内（啮齿动物）研究中的毒性。近年来，使用斑马鱼（Danio rerio）来研究化学品，病原体和损伤的炎症反应大大增加。斑马鱼也被用于研究炎症在疾病发病机制中的作用和药物发现。在免疫细胞（例如巨噬细胞和嗜中性粒细胞）中表达荧光蛋白的转基因菌株的可用性使得能够可视化和定量感兴趣的靶位点中的免疫细胞积累。因此，我们建议，转基因斑马鱼有很大的效用，通过调查的炎症反应筛选NM的毒性。事实上，我们已经成功地使用了非保护的生命阶段的转基因斑马鱼与荧光中性粒细胞（Tg（mpx：EGFP 114），以研究炎症反应的NM。在纳米毒理学中更广泛地使用转基因斑马鱼可以减少对啮齿动物的依赖，从而加强3Rs原则的实施。随着斑马鱼越来越受欢迎，为新用户提供使用指导是及时的。在这里，我们将反思：这些问题包括：可用于模拟人类/啮齿动物接触的接触途径、哪些转基因品系和生命阶段最适合研究炎症反应、斑马鱼胚胎/幼体的选择标准、适当对照的纳入、剂量选择和样本大小的重要性以及如何量化（炎症）反应。希望我们的建议将支持标准协议的发展，可用于评估NM是否激活炎症反应。重要的是，讨论的主题不仅限于纳米材料，而且还与斑马鱼在生态毒理学或人类健康研究中的应用有关。
Despite the increasing exploitation of nanomaterials (NMs) in an array of consumer products, there are uncertainties regarding their potential adverse impact on human health. Investigation of whether NMs activate a pro-inflammatory response is routinely used to assess their toxicity in in vitro and in vivo (rodent) studies. The use of zebrafish (Danio rerio) to investigate inflammatory responses to chemicals, pathogens and injury has increased considerably over recent years. Zebrafish have also been used to investigate the role of inflammation in disease pathogenesis and for drug discovery. Availability of transgenic strains which express fluorescent proteins in immune cells (e.g. macrophages and neutrophils) enables the visualization and quantification of immune cell accumulation in the target site(s) of interest. We therefore propose that transgenic zebrafish have great utility for screening the toxicity of NMs via investigation of inflammatory responses. Indeed, we have successfully used non-protected life stages of transgenic zebrafish with fluorescent neutrophils (Tg(mpx:EGFP114) to investigate inflammatory responses to NMs. The more widespread use of transgenic zebrafish in nanotoxicology could reduce the reliance placed on rodents and thereby enhance the implementation of the 3Rs principles. As zebrafish continue to grow in popularity it is timely to offer guidance to new users on their use. Here we will reflect on: exposure routes that can adopted to mimic human/rodent exposure, what transgenic strains and life stages are best suited to investigate inflammatory responses, selection criteria for zebrafish embryos/larvae, the inclusion of appropriate controls, the importance of dose selection and sample size, and how the (inflammatory) response can be quantified. It is hoped that our recommendations will support the development of standard protocols that can be used to assess whether NMs activate inflammatory responses. Importantly, the themes discussed are not restricted to NMs but relevant also to zebrafish application in ecotoxicology or human health focused studies.