Zebrafish 24 hpf-embryos as a Model Organism for Measuring Oxygen Consumption Rate (OCR).

IF 3.2 4区医学 Q1 Pharmacology, Toxicology and Pharmaceutics

Toxicology Mechanisms and Methods Pub Date : 2025-07-23 DOI:10.1080/15376516.2025.2537894

P V L Peixoto, C C da Silva, L C Pereira

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引用次数: 0

Abstract

The alterations in mitochondrial function are involved in various pathological conditions and hence the evaluation of such damage is crucial to determine the mitotoxic potential of different chemicals. Due to their 71% genetic similarity to humans, the similar mitochondrial functions, and metabolic processes of zebrafish (Danio rerio) have been used in analyses for this purpose. The aim of this study was to establish a standardized technique to assess mitochondrial dysfunction by analyzing oxygen consumption rate (OCR) in zebrafish embryos 24 hours post-fertilization using the OROBOROS O2k Oxygraph. The technique involved the use of mitochondrial modulators and the OROBOROS O2k Oxygraph to directly assess mitochondrial and electron transport chain complex activity during embryonic development. Embryos were treated with a respiratory medium supplemented with malate, succinate, and pyruvate, and with digitonin to permeabilize the chorion and membranes for mitochondrial analysis. OCR measurements were performed in the presence of specific mitochondrial modulators: oligomycin, FCCP, rotenone and antimycin A. Optimized evaluation was achieved using 20 embryos per assay. Therefore, through the development of a protocol for synchronization analysis of OCR in zebrafish embryos, several parameters related to the effectiveness of the oxidative phosphorylation process could be rapidly determined. Since zebrafish are particularly useful to study mitochondrial dysfunction in toxicants, this protocol describes the procedure to quantitate the effect of toxicants on mitochondrial activity which turns out to be valuable to understanding the mechanism of xenobiotic toxicity.

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斑马鱼24个hpf胚胎作为测量耗氧量（OCR）的模式生物。

线粒体功能的改变涉及各种病理条件，因此评估这种损害对于确定不同化学物质的有丝分裂毒性潜力至关重要。由于它们与人类71%的遗传相似性，斑马鱼（Danio rerio）相似的线粒体功能和代谢过程已被用于这一目的的分析。本研究的目的是建立一种标准化的技术，通过使用OROBOROS O2k氧图分析受精后24小时斑马鱼胚胎的耗氧量（OCR）来评估线粒体功能障碍。该技术包括使用线粒体调节剂和OROBOROS O2k氧合仪来直接评估胚胎发育过程中线粒体和电子传递链复合物的活性。胚胎用含有苹果酸盐、琥珀酸盐和丙酮酸盐的呼吸培养基和洋地黄苷处理，以渗透绒毛膜和膜，进行线粒体分析。在特定线粒体调节剂（寡霉素、FCCP、鱼藤酮和抗霉素a）存在的情况下进行OCR测量。每次检测使用20个胚胎进行优化评估。因此，通过制定斑马鱼胚胎OCR同步分析方案，可以快速确定与氧化磷酸化过程有效性相关的几个参数。由于斑马鱼对研究毒物中的线粒体功能障碍特别有用，本方案描述了定量毒物对线粒体活性影响的程序，这对理解外源毒性的机制是有价值的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Toxicology Mechanisms and Methods 医学-毒理学

CiteScore

6.60

自引率

3.10%

发文量

审稿时长

6-12 weeks

期刊介绍： Toxicology Mechanisms and Methods is a peer-reviewed journal whose aim is twofold. Firstly, the journal contains original research on subjects dealing with the mechanisms by which foreign chemicals cause toxic tissue injury. Chemical substances of interest include industrial compounds, environmental pollutants, hazardous wastes, drugs, pesticides, and chemical warfare agents. The scope of the journal spans from molecular and cellular mechanisms of action to the consideration of mechanistic evidence in establishing regulatory policy. Secondly, the journal addresses aspects of the development, validation, and application of new and existing laboratory methods, techniques, and equipment.