Da'as Sahar I, Balci Tugce B, Berman Jason N
IWK Health Centre, Dalhousie University, 9700, 5850/5980 University Avenue, Halifax, NS, Canada, B3K 6R8.
Methods Mol Biol. 2015;1220:29-57. doi: 10.1007/978-1-4939-1568-2_3.
The many advantages of the zebrafish model provide a unique opportunity to integrate the tools of developmental embryology, transgenesis, and functional assays to elucidate the molecular pathways underlying hematopoiesis and for modeling human blood diseases. These methodologies have recently been applied to the zebrafish mast cell lineage and have resulted in a better understanding of vertebrate mast cell biology. By employing whole-mount in situ hybridization alone and in combination with co-localization approaches, fluorescence-activated cell sorting (FACS), and morpholino gene knockdown studies, new insights into early mast cell transcriptional regulation and ontogeny have been exposed in vivo. Transgenic strategies have permitted the modeling of human mast cell diseases, like systemic mastocytosis in zebrafish, which can subsequently be exploited for high-throughput chemical screens to identify potential therapies in these conditions. Mast cell functional assays have been adapted to zebrafish providing the opportunity to utilize this model for interrogating the cellular players in innate and adaptive immunity and as a live animal readout for drug responses in allergic and inflammatory reactions. These techniques are detailed in the following chapter.
斑马鱼模型的诸多优势提供了一个独特的机会，可整合发育胚胎学、转基因技术和功能分析工具，以阐明造血作用背后的分子途径，并用于模拟人类血液疾病。这些方法最近已应用于斑马鱼肥大细胞谱系，从而对脊椎动物肥大细胞生物学有了更好的理解。通过单独使用全胚胎原位杂交，并结合共定位方法、荧光激活细胞分选（FACS）和吗啉代基因敲低研究，在体内揭示了对早期肥大细胞转录调控和个体发生的新见解。转基因策略已允许对人类肥大细胞疾病进行建模，如斑马鱼中的系统性肥大细胞增多症，随后可用于高通量化学筛选，以确定这些病症的潜在疗法。肥大细胞功能分析已适用于斑马鱼，这为利用该模型研究先天免疫和适应性免疫中的细胞参与者以及作为过敏和炎症反应中药物反应的活体动物读数提供了机会。这些技术将在以下章节中详细介绍。
