Weaver Cory J, Leung Yuk Fai, Suter Daniel M
Department of Biological Sciences, Purdue University, West Lafayette, Indiana, 47907.
Bindley Bioscience Center, Purdue University, West Lafayette, Indiana, 47907.
J Comp Neurol. 2016 Jul 1;524(10):2130-41. doi: 10.1002/cne.23938. Epub 2015 Dec 29.
Nicotinamide dinucleotide phosphate oxidases (NOX) control various cellular signaling cascades. In the nervous system, there is recent evidence that NOX-derived reactive oxygen species (ROS) regulate neurite outgrowth, regeneration, and stem cell proliferation; however, a comprehensive NOX gene expression analysis is missing for all major model systems. Zebrafish embryos provide an excellent model system to study neurodevelopment and regeneration because they develop quickly and are well suited for in vivo imaging and molecular approaches. Although the sequences of five NOX genes (nox1, nox2/cybb, nox4, nox5, and duox) have been identified in the zebrafish genome, nothing is known about their expression pattern. Here, we used quantitative polymerase chain reaction combined with in situ hybridization to develop a catalog of nox1, nox2/cybb, nox5, and duox expression in zebrafish during early nervous system development from 12 to 48 hours post fertilization. We found that expression levels of nox1, nox5, and duox are dynamic during the first 2 days of development, whereas nox2/cybb levels remain remarkably stable. By sectioning in situ hybridized embryos, we found a pattern of broad and overlapping NOX isoform expression at 1 and 1.5 days post fertilization. After 2 days of development, a few brain regions displayed increased NOX expression levels. Collectively, these results represent the first comprehensive analysis of NOX gene expression in the zebrafish and will provide a basis for future studies aimed at determining the functions of NOX enzymes in neurodevelopment and regeneration. J. Comp. Neurol. 524:2130-2141, 2016. © 2015 Wiley Periodicals, Inc.
烟酰胺腺嘌呤二核苷酸磷酸氧化酶（NOX）控制着各种细胞信号级联反应。在神经系统中，最近有证据表明，NOX衍生的活性氧（ROS）调节神经突生长、再生和干细胞增殖；然而，所有主要模型系统都缺乏全面的NOX基因表达分析。斑马鱼胚胎为研究神经发育和再生提供了一个极好的模型系统，因为它们发育迅速，非常适合体内成像和分子方法。虽然在斑马鱼基因组中已经鉴定出五个NOX基因（nox1、nox2/cybb、nox4、nox5和duox）的序列，但对它们的表达模式却一无所知。在这里，我们使用定量聚合酶链反应结合原位杂交技术，建立了受精后12至48小时斑马鱼早期神经系统发育过程中nox1、nox2/cybb、nox5和duox表达的目录。我们发现，在发育的前两天，nox1、nox5和duox的表达水平是动态的，而nox2/cybb的水平则保持显著稳定。通过对原位杂交胚胎进行切片，我们发现在受精后1天和1.5天，NOX异构体表达呈现广泛且重叠的模式。发育2天后，一些脑区的NOX表达水平升高。总的来说，这些结果代表了对斑马鱼NOX基因表达的首次全面分析，并将为未来旨在确定NOX酶在神经发育和再生中的功能的研究提供基础。《比较神经学杂志》524:2130 - 2141, 2016。© 2015威利期刊公司。
