Optimization of MSI technologies for environmental toxicology: A case study with Zebrafish eleutheroembryos
The Zebrafish (Danio rerio) has become a powerful model organism in a wide range of scientific fields, including ecotoxicology. The Zebrafish model presents various advantages concerning other common model organisms such as its effortless manipulation or large offspring [ref]. Strikingly, toxicological data from zebrafish can be extrapolated not only to aquatic species but also to other vertebrates, including humans. Moreover, zebrafish embryos are considered an excellent alternative animal model with fewer ethical restrictions, ensuring the fulfillment of the 3R’s principle (Replacement, Reduction, and Refinement) in animal research (. For that reason, their use in ecotoxicological studies and, particularly, in endocrine-disrupting chemicals (EDCs) assessment is broadly boosted. Bulk omic technologies have contributed to environmental toxicology to deepen an organism's response to pollutants at the molecular level. However, reporting the molecular information of individual cell types in addition to their spatial organization is unachievable. In heterogeneous samples such as a whole animal or embryo, it is critical to improve the knowledge in the mode of action of pollutants be capable of outlining these information. To overcome these challenges, breakthrough technologies have emerged to encompass single-cell and spatially resolved omics, including mass spectrometry imaging (MSI). Particularly relevant is the use of Matrix-Assisted Laser Desorption/Ionization (MALDI-MSI) owing to it providing a favorable balance between sample preparation, chemical sensitivity, and spatial resolution. Despite its outstanding features, MALDI approaches have some limitations in lipidomics studies. For instance, the conditions for the optimal ionization of certain lipid classes (i.e., sterols) or the spatial resolution compared to other MSI techniques. For that reason, different analytical techniques (e.g., on-tissue-derivatization, and metal deposition) have recently been developed to improve MALDI capabilities. Also, the state–of–of–the–art laser-post ionization coupled with the MALDI (MALDI-2) tool has emerged as a potential new game-changer in the MSI field. MALDI-2 improve sensitivity for a high number of suppressed lipid classes leading to a decrease in pixel size. In this study, we developed and optimized a spatial lipidomics protocol to analyze zebrafish embryo sections using both MALDI and MALDI-2-MSI with a lateral spatial resolution of up to 5 μm. Our results revealed the presence of different lipid clusters corresponding to different sections of the zebrafish embryo. Therefore, these results demonstrate the usefulness of spatial omics studies in this biological model, particularly underlining possible lipid biomarkers for relevant tissues such as eye or brain.
| Main Authors: | , , , , , , |
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| Format: | comunicación de congreso biblioteca |
| Language: | English |
| Published: |
2023
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| Subjects: | Environmental toxicology, Ensure availability and sustainable management of water and sanitation for all, |
| Online Access: | http://hdl.handle.net/10261/355726 |
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