Mass Spectrometry-Based Zebrafish Toxicometabolomics: A Review of Analytical and Data Quality Challenges

Abstract

Metabolomics has achieved great progress over the last 20 years, and it is currently considered a mature research field. As a result, the number of applications in toxicology, biomarker, and drug discovery has also increased. Toxicometabolomics has emerged as a powerful strategy to provide complementary information to study molecular-level toxic effects, which can be combined with a wide range of toxicological assessments and models. The zebrafish model has gained importance in recent decades as a bridging tool between in vitro assays and mammalian in vivo studies in the field of toxicology. Furthermore, as this vertebrate model is a low-cost system and features highly conserved metabolic pathways found in humans and mammalian models, it is a promising tool for toxicometabolomics. This short review aims to introduce zebrafish researchers interested in understanding the effects of chemical exposure using metabolomics to the challenges and possibilities of the field, with a special focus on toxicometabolomics-based mass spectrometry. The overall goal is to provide insights into analytical strategies to generate and identify high-quality metabolomic experiments focusing on quality management systems (QMS) and the importance of data reporting and sharing.

Lipidomics profiling of zebrafish liver through untargeted liquid chromatography‐high resolution mass spectrometry

“…Although untargeted methods aim to cover as many metabolites as possible, LC-HRMS methods are limited to certain metabolite classes (e.g., lipid mediators such as oxylipins require specific sample treatment and high instrumental sensitivity [40]) [1]. Due to the structural diversity of lipids, different studies and analytical platforms should be combined to reveal the most accurate composition of a specific organism or tissue.…”

“…Apart from the lack of a stomach, the zebrafish digestive system is highly similar to that of mammals including humans, both structurally and functionally. Metabolic functions and processes are also highly conserved, including the major metabolic pathways and hormones regulating e.g., digestion, appetite, and glucose homeostasis [1,2]. Zebrafish are increasingly being used to study the effects of so-called metabolic disruptors, a class of endocrine-disrupting compounds that increase the susceptibility to metabolic disorders.…”

“…The most commonly used lipidomics strategy consists of extraction using organic solvents, followed by reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) [5,7]. Untargeted high-resolution MS (HRMS) methods combined with liquid-liquid extraction (LLE) provide comprehensive coverage of multiple lipid classes [1,8]. In many applications, sample preparation methods developed for a specific sample type will be applied to a different matrix.…”

“…Model organisms play a significant role in understanding the functions of lipids. Qualitative information on lipid composition that can be used for further quantitative measurements to study metabolic diseases and exposure to toxicants is of special interest since this information is still scarce [1]. In this study, an analytical workflow for zebrafish liver tissues was explored using an untargeted RPLC-HRMS-based platform previously applied for liver cell extracts [15].…”

Untargeted Metabolomics Reveals Lipid Impairment in the Liver of Adult Zebrafish (Danio rerio) Exposed to Carbendazim

“…These data indicate the need for further studies on the effects of carbendazim exposure. In this regard, the zebrafish ( Danio rerio ) is a medium‐ to high‐throughput vertebrate toxicological model that is routinely used to provide relevant information regarding the acute and long‐term effects of xenobiotics (Silva et al, 2021). Thus, zebrafish is a prominent species for evaluations of the effects of exposure to environmental contaminants (Almeida et al, 2021; Loncar & Smital, 2018; Lu et al, 2016).…”