A high-throughput chemically induced inflammation assay in zebrafish.
A high-throughput chemically induced inflammation assay in zebrafish.
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DOI：
10.1186/1741-7007-8-151
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发表时间：
2010-12-22
期刊：
影响因子：
5.4
通讯作者：
Allende ML
中科院分区：
文献类型：
--
作者：
d'Alençon CA;Peña OA;Wittmann C;Gallardo VE;Jones RA;Loosli F;Liebel U;Grabher C;Allende ML
对先天免疫的研究得益于斑马鱼作为模型系统的引入。在白细胞群体中表达荧光蛋白的转基因鱼允许直接、定量地可视化体内炎症反应。已经提出，该动物模型可用于高通量筛选，旨在鉴定新型免疫调节先导化合物。然而，目前的检测需要对鱼进行单独的侵入性操作，从而阻止了高含量筛选。在这里，我们表明，特定的，非侵入性的损伤侧线神经肥大细胞可以诱导一个强大的急性炎症反应。暴露于亚致死浓度的硫酸铜的鱼苗选择性损害的感觉毛细胞群诱导浸润的白细胞在20分钟内的神经瘤。炎症可以使用在白细胞中表达荧光蛋白的转基因鱼或通过在固定的幼虫中的组织化学测定来真实的时间测定。我们证明了这种方法对于化学和遗传筛查的有用性，以检测免疫调节化合物和影响白细胞反应的突变的影响。此外，我们通过使用定制的自动成像和处理系统将该测定转化为高通量筛选方法，该系统可以量化炎症反应的程度。这种方法可以快速筛选数千种化合物或经突变的斑马鱼对炎症的影响，并能够识别调节先天免疫的新参与者和新免疫调节疗法的潜在先导化合物。我们将这种方法称为化学诱导炎症测定或ChIn测定。参见评论文章：http://www.biomedcentral.com/1741-7007/8/148。
Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening. Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction. This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay. See Commentary article: http://www.biomedcentral.com/1741-7007/8/148.
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影响因子：
4.5
作者：
Behra M;Bradsher J;Sougrat R;Gallardo V;Allende ML;Burgess SM
通讯作者：
Burgess SM
影响因子：
2.9
作者：
Mathias, Jonathan R.;Dodd, M. Ernest;Walters, Kevin B.;Yoo, Sa Kan;Ranheim, Erik A.;Huttenlocher, Anna
通讯作者：
Huttenlocher, Anna
影响因子：
4
作者：
Mathias, Jonathan R.;Dodd, M. Ernest;Huttenlocher, Anna
通讯作者：
Huttenlocher, Anna
影响因子：
3.9
作者：
Martin, Jane S.;Renshaw, Stephen A.
通讯作者：
Renshaw, Stephen A.
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF