PUBLICATION
High-fat Feeding Paradigm for Larval Zebrafish: Feeding, Live Imaging, and Quantification of Food Intake
- Authors
- Otis, J.P., Farber, S.A.
- ID
- ZDB-PUB-161115-3
- Date
- 2016
- Source
- Journal of visualized experiments : JoVE (116): (Journal)
- Registered Authors
- Farber, Steven, Otis, Jessica P.
- Keywords
- none
- MeSH Terms
-
- Dietary Fats*
- Egg Yolk
- Animals
- Chickens
- Zebrafish*
- Animal Feed*
- Larva
- Fatty Acids
- PubMed
- 27842350 Full text @ J. Vis. Exp.
Citation
Otis, J.P., Farber, S.A. (2016) High-fat Feeding Paradigm for Larval Zebrafish: Feeding, Live Imaging, and Quantification of Food Intake. Journal of visualized experiments : JoVE. (116).
Abstract
Zebrafish are emerging as a model of dietary lipid processing and metabolic disease. This protocol describes how to feed larval zebrafish a lipid-rich meal, which consists of an emulsion of chicken egg yolk liposomes created by sonicating egg yolk in embryo media. Detailed instructions are provided to screen larvae for egg yolk consumption so that larvae that fail to feed will not confound experimental results. The chicken egg yolk liposomes can be spiked with fluorescent lipid analogs, including fatty acids and cholesterol, enabling both systemic and subcellular visualization of dietary lipid processing. Several methods are described to mount larvae that are conducive to short- and long-term live imaging with both upright and inverted objectives at high and low magnification. Additionally presented is an assay to quantify larval food intake by extracting the lipids of larvae fed fluorescent lipid analogs, spotting the lipids on a thin layer chromatography plate, and quantifying the fluorescence. Finally, critical aspects of the procedures, important controls, options for modifying the protocols to address specific experimental questions, and potential limitations are discussed. These techniques can be applied not only to focused, hypothesis driven inquiries, but also to a variety of screens and live imaging techniques to study dietary lipid metabolism and the control of food intake.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping