title: Bioenergetic profiling of zebrafish embryonic development.
abstract: Many debilitating conditions are linked to bioenergetic defects. Developing screens to probe the genetic and/or chemical basis for such links has proved intractable. Furthermore, there is a need for a physiologically relevant assay of bioenergetics in whole organisms, especially for early stages in life where perturbations could increase disease susceptibility with aging. Thus, we asked whether we could screen bioenergetics and mitochondrial function in the developing zebrafish embryo. We present a multiplexed method to assay bioenergetics in zebrafish embryos from the blastula period (3 hours post-fertilization, hpf) through to hatching (48 hpf). In proof of principle experiments, we measured respiration and acid extrusion of developing zebrafish embryos. We quantified respiratory coupling to various bioenergetic functions by using specific pharmacological inhibitors of bioenergetic pathways. We demonstrate that changes in the coupling to ATP turnover and proton leak are correlated with developmental stage. The multiwell format of this assay enables the user to screen for the effects of drugs and environmental agents on bioenergetics in the zebrafish embryo with high sensitivity and reproducibility.
authors: Department of Pharmaceutical and Biomedical Sciences, Medical University of South Carolina, Charleston, South Carolina, United States of America.
authors: Krista D
authors: KD
authors: Stackley
authors: Craig C
authors: CC
authors: Beeson
authors: Jennifer J
authors: JJ
authors: Rahn
authors: Sherine S L
authors: SS
authors: Chan
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D019069:Cell Respiration / Q000187:drug effects; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004734:Energy Metabolism / Q000187:drug effects; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D046228:Microarray Analysis / Q000379:methods*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D011522:Protons; D015203:Reproducibility of Results; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism
chemical_list: D011522:Protons; D000255:Adenosine Triphosphate