Bioenergetic profiling of zebrafish embryonic development.
Bioenergetic profiling of zebrafish embryonic development.
复制标题
DOI：
10.1371/journal.pone.0025652
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Chan SS
中科院分区：
文献类型：
--
作者：
Stackley KD;Beeson CC;Rahn JJ;Chan SS
许多使人衰弱的疾病都与生物能量缺陷有关。开发筛选来探测这些联系的遗传和/或化学基础已被证明是棘手的。此外，有必要对整个生物体的生物能量学进行生理相关的测定，特别是在生命的早期阶段，随着年龄的增长，扰动可能会增加疾病的易感性。因此，我们想知道我们是否可以在发育中的斑马鱼胚胎中筛选生物能量学和线粒体功能。我们提出了一种多重方法来测定斑马鱼胚胎从囊胚期（受精后3小时，hpf）到孵化（48 hpf）的生物能量学。在证明原理的实验中，我们测量了发育中的斑马鱼胚胎的呼吸和酸挤压。我们通过使用生物能量途径的特定药理抑制剂来量化呼吸耦合到各种生物能量功能。我们证明了ATP转换和质子泄漏耦合的变化与发育阶段有关。该检测的多孔格式使用户能够筛选药物和环境因子对斑马鱼胚胎生物能量学的影响，具有高灵敏度和可重复性。
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.
登录
查看更多内容
影响因子：
13.8
作者：
Azzu, Vian;Brand, Martin D.
通讯作者：
Brand, Martin D.
影响因子：
7.4
作者：
Gerencser, Akos A.;Neilson, Andy;Choi, Sung W.;Edman, Ursula;Yadava, Nagendra;Oh, Richard J.;Ferrick, David A.;Nicholls, David G.;Brand, Martin D.
通讯作者：
Brand, Martin D.
影响因子：
2
作者：
Bang, A;Gronkjær, P;Malte, H
通讯作者：
Malte, H
影响因子：
4.1
作者：
BRAND, MD;COUTURE, P;HULBERT, AJ
通讯作者：
HULBERT, AJ
影响因子：
2.7
作者：
Brownlie, A;Hersey, C;Zon, L
通讯作者：
Zon, L