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Metabolic profile analysis of zebrafish embryos
journal contribution
posted on 2022-09-29, 03:19 authored by Y Gibert, Sean McgeeSean Mcgee, Alister WardAlister WardA growing goal in the field of metabolism is to determine the impact of genetics on different aspects of mitochondrial function. Understanding these relationships will help to understand the underlying etiology for a range of diseases linked with mitochondrial dysfunction, such as diabetes and obesity. Recent advances in instrumentation, has enabled the monitoring of distinct parameters of mitochondrial function in cell lines or tissue explants. Here we present a method for a rapid and sensitive analysis of mitochondrial function parameters in vivo during zebrafish embryonic development using the Seahorse bioscience XF 24 extracellular flux analyser. This protocol utilizes the Islet Capture microplates where a single embryo is placed in each well, allowing measurement of bioenergetics, including: (i) basal respiration; (ii) basal mitochondrial respiration (iii) mitochondrial respiration due to ATP turnover; (iv) mitochondrial uncoupled respiration or proton leak and (iv) maximum respiration. Using this approach embryonic zebrafish respiration parameters can be compared between wild type and genetically altered embryos (mutant, gene over-expression or gene knockdown) or those manipulated pharmacologically. It is anticipated that dissemination of this protocol will provide researchers with new tools to analyse the genetic basis of metabolic disorders in vivo in this relevant vertebrate animal model.
History
Journal
Journal of visualized experimentsVolume
71Issue
71Season
Article e4300Article number
ARTN e4300Pagination
1-5Publisher
Journal of Visualized ExperimentsLocation
Cambridge, Mass.Publisher DOI
ISSN
1940-087XeISSN
1940-087XLanguage
engCopyright notice
2013, JoVEUsage metrics
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Keywords
Science & TechnologyMultidisciplinary SciencesScience & Technology - Other TopicsDevelopmental BiologyIssue 71GeneticsBiochemistryCellular BiologyMolecular BiologyPhysiologyEmbryologyMetabolismMetabolomicsmetabolic profilerespirationmitochondriaATPdevelopmentOil Red O stainingzebrafishDanio rerioanimal modelAdenosine TriphosphateAnimalsEnergy MetabolismMitochondriaZebrafishDiabetesMetabolic and endocrineGeneric health relevanceNHMRC 1030474119999 Medical and Health Sciences not elsewhere classifiedC1 Refereed article in a scholarly journal920104 DiabetesC Journal articleSchool of MedicineCognitive Sciences