Whole-body metabolism is regulated by a complex interplay between multiple organs. The adult zebrafish is an attractive model to study such processes because it has all of the organs that are required for metabolic control in humans. Its relatively small size and low blood volume, however, makes metabolomics analysis of single animals technically challenging. Here, we tested multiple dissection and quenching techniques for performing metabolomics analysis of single adult zebrafish on the basis of precision and accuracy. Additionally, we tested three different strategies for introducing isotope tracers: direct addition to the tank water, oral gavage, and intraperitoneal injection. Each was evaluated according to the total isotopic enrichment achieved and the ability to reach isotopic steady state. An example application in which the optimized methods are applied to study systemic changes in metabolism as a result of melanoma will be presented.

Developing a metabolomics platform for isotope-tracer analysis in adult zebrafish

ACS Fall 2021

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