Abstract
The zebrafish model facilitates the study of lipid metabolism and transport during development. Here, we outline methods to introduce traceable fluorescent or radiolabeled fatty acids into zebrafish embryos and larvae at various developmental stages. Labeled fatty acids can be injected into the large yolk cell prior to the development of digestive organs when the larvae is entirely dependent on the yolk for its nutrition (lecithotrophic state). Once zebrafish are able to consume exogenous food, labeled fatty acids can be incorporated into their food. Our group and others have demonstrated that the transport and processing of these injected or ingested fatty acid analogs can be followed through microscopy and/or biochemical analysis. These techniques can be easily combined with targeted antisense approaches, transgenics, or drug treatments (see Note 1 ), allowing studies of lipid cell biology and metabolism that are exceedingly difficult or impossible in mammals.
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Acknowledgments
The authors would like to thank Drs. James Walters and Juliana Carten for the initial development of the fluorescent liposome feeding assay; Dr. Rosa Miyares and Jennifer Anderson for the initial development of the oil injection technique; Drs. Juliana Carten, Rosa Miyares, and Vitor deRezende for the initial exploration into the use of radiolabeled fatty acids and lipids in zebrafish metabolic assays; and Mahmud Siddiqi for his technical support with microscopy. This work was supported in part by the National Institute of Diabetes and Digestive and Kidney (NIDDK) [grant numbers RO1DK093399 to S.A.F., RO1GM63904 to The Zebrafish Functional Genomics Consortium (Stephen Ekker and S.A.F.)]. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health (NIH). Additional support for this work was provided by the Carnegie Institution for Science endowment and the G. Harold and Leila Y. Mathers Charitable Foundation to the laboratory of S.A.F.
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Zeituni, E.M., Farber, S.A. (2016). Studying Lipid Metabolism and Transport During Zebrafish Development. In: Kawakami, K., Patton, E., Orger, M. (eds) Zebrafish. Methods in Molecular Biology, vol 1451. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3771-4_16
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DOI: https://doi.org/10.1007/978-1-4939-3771-4_16
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