DanioLabs, Cambridge, UK.
Inflamm Bowel Dis. 2010 Jul;16(7):1162-72. doi: 10.1002/ibd.21200.
The aim of this study was to develop a model of inflammatory bowel disease (IBD) in zebrafish larvae, together with a method for the rapid assessment of gut morphology and function in vivo thereby enabling medium-throughput compound screening.
Assays were performed using larval zebrafish from 3-8 days postfertilization (d.p.f.) in 96-well plates. Gut morphology and peristalsis were observed in vivo using fluorescent imaging following ingestion of fluorescent dyes. IBD was induced by addition of 2,4,6-trinitrobenzenesulfonic acid (TNBS) to the medium within the well. Pathology was assessed in vivo using fluorescent imaging and postmortem by histology, immunohistochemistry, and electron microscopy. Therapeutic compounds were evaluated by coadministration with TNBS.
A novel method of investigating gut architecture and peristalsis was devised using fluorescent imaging of live zebrafish larvae. Archetypal changes in gut architecture consistent with colitis were observed throughout the gut. Significant changes in goblet cell number and tumor necrosis factor alpha (TNF-alpha) antibody staining were used to quantify disease severity and rescue. Prednisolone and 5-amino salicylic acid treatment ameliorated the disease changes. Candidate therapeutic compounds (NOS inhibitors, thalidomide, and parthenolide) were assessed and a dissociation was observed between efficacy assessed using a single biochemical measure (TNF-alpha staining) versus an assessment of the entire disease state.
Gut physiology and pathology relevant to human disease state can be rapidly modeled in zebrafish larvae. The model is suitable for medium-throughput chemical screens and is amenable to genetic manipulation, hence offers a powerful novel premammalian adjunct to the study of gastrointestinal disease.
本研究旨在建立斑马鱼幼鱼炎症性肠病（IBD）模型，同时开发一种快速评估肠道形态和功能的方法，从而实现高通量化合物筛选。
在 96 孔板中，对 3-8 日龄（d.p.f.）的斑马鱼幼鱼进行检测。通过荧光染料摄取后，活体荧光成像观察肠道形态和蠕动。在孔内培养基中加入 2,4,6-三硝基苯磺酸（TNBS）可诱导 IBD。通过活体荧光成像和死后组织学、免疫组织化学和电子显微镜检查评估体内病理学。通过与 TNBS 共给药评估治疗性化合物。
开发了一种新的方法，通过活体斑马鱼幼鱼的荧光成像研究肠道结构和蠕动。观察到整个肠道出现与结肠炎一致的典型肠道结构变化。杯状细胞数量和肿瘤坏死因子-α（TNF-α）抗体染色的显著变化用于量化疾病严重程度和恢复情况。泼尼松龙和 5-氨基水杨酸治疗可改善疾病变化。评估候选治疗性化合物（NOS 抑制剂、沙利度胺和白头翁内酯）时，发现单一生化指标（TNF-α染色）评估的疗效与整个疾病状态评估之间存在差异。
与人类疾病状态相关的肠道生理学和病理学可在斑马鱼幼鱼中快速建模。该模型适用于高通量化学筛选，并且易于进行遗传操作，因此为胃肠道疾病的研究提供了一种强大的新型哺乳动物前体。
