bioenergetic profiling of zebrafish embryonic development斑马鱼胚胎发育的生物能量学分析.pdf

Bioenergetic Profiling of Zebrafish Embryonic Development Krista D. Stackley, Craig C. Beeson, Jennifer J. Rahn, Sherine S. L. Chan* Department of Pharmaceutical and Biomedical Sciences, Medical University of South Carolina, Charleston, South Carolina, United States of America 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. Citation: Stackley KD, Beeson CC, Rahn JJ, Chan SSL (2011) Bioenergetic Profiling of Zebrafish Embryonic Development. PLoS ONE 6(9): e25652. doi:10.1371/ journal.pone.0025652 ´ ˜ Editor: Alicia J. Kowaltowski, Instituto de Quımica - Uni