Jung H M, Isogai S, Kamei M, Castranova D, Gore A V, Weinstein B M
National Institute of Child Health and Human Development, Bethesda, MD, United States.
Iwate Medical University, Morioka, Japan.
Methods Cell Biol. 2016;133:69-103. doi: 10.1016/bs.mcb.2016.03.023. Epub 2016 Apr 18.
Blood vessels supply tissues and organs with oxygen, nutrients, cellular, and humoral factors, while lymphatic vessels regulate tissue fluid homeostasis, immune trafficking, and dietary fat absorption. Understanding the mechanisms of vascular morphogenesis has become a subject of intense clinical interest because of the close association of both types of vessels with pathogenesis of a broad spectrum of human diseases. The zebrafish provides a powerful animal model to study vascular morphogenesis because of their small, accessible, and transparent embryos. These unique features of zebrafish embryos permit sophisticated high-resolution live imaging of even deeply localized vessels during embryonic development and even in adult tissues. In this chapter, we summarize various methods for blood and lymphatic vessel imaging in zebrafish, including nonvital resin injection-based or dye injection-based vessel visualization, and alkaline phosphatase staining. We also provide protocols for vital imaging of vessels using microangiography or transgenic fluorescent reporter zebrafish lines.
血管为组织和器官提供氧气、营养物质、细胞和体液因子,而淋巴管则调节组织液稳态、免疫细胞运输和膳食脂肪吸收。由于这两种血管与多种人类疾病的发病机制密切相关,了解血管形态发生的机制已成为临床研究的热点。斑马鱼因其胚胎小、易于操作且透明,为研究血管形态发生提供了一个强大的动物模型。斑马鱼胚胎的这些独特特征使得在胚胎发育期间甚至在成体组织中,即使是对深部定位的血管也能进行复杂的高分辨率实时成像。在本章中,我们总结了斑马鱼血液和淋巴管成像的各种方法,包括基于非活体树脂注射或染料注射的血管可视化方法,以及碱性磷酸酶染色。我们还提供了使用微血管造影或转基因荧光报告斑马鱼品系对血管进行活体成像的方案。