Zebrafish model of the blood-brain barrier: morphological and permeability studies.
Zebrafish model of the blood-brain barrier: morphological and permeability studies.
复制标题
DOI：
10.1007/978-1-60761-938-3_18
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Baird, Andrew
中科院分区：
文献类型：
--
作者：
Eliceiri, Brian P;Gonzalez, Ana Maria;Baird, Andrew
血脑屏障（BBB）是单层内皮细胞，其由独特的基底膜的邻近性和包括周细胞、星形胶质细胞和神经元的专门细胞亚群之间的严格控制的分子相互作用来调节。这些细胞一起工作，形成一个神经血管单位（NVU），致力于局部调节大脑中的血管功能和BBB的完整性。因此，NVU的细胞-基质-细胞相互作用的内在复杂性使得分析细胞培养物、组织外植体、甚至动物模型中的基因功能变得困难，并且不能在体内研究BBB中的基因功能已经成为推进BBB研究的关键障碍。斑马鱼已经成为一种主要的脊椎动物生物体，可以模拟和分析体内复杂的细胞相互作用和胚胎发育的遗传机制。为此，我们提供了一个技术概述的程序，可用于在斑马鱼BBB的完整性分析，重点是对成年鱼的脑血管系统的BBB现在被定义。用于测量BBB的功能完整性、细胞生物学和超微结构的技术分别包括渗透性测定、报告基因的荧光成像和电子显微镜。每一种都可以应用于突变鱼的功能分析，以表征损害BBB完整性的病理损伤的分子后遗症的方式。由于与高等脊椎动物相比，斑马鱼的遗传学和细胞生物学具有高度保守性，因此可以在斑马鱼模型中使用药物发现技术，以补充其他模型系统中的药物开发研究。
The blood-brain barrier (BBB) is a monolayer of endothelial cells that is regulated by the proximity of a unique basement membrane and a tightly controlled molecular interaction between specialized subsets of cells including pericytes, astrocytes, and neurons. Working together, these cells form a neurovascular unit (NVU) that is dedicated to the local regulation of vascular function in the brain and BBB integrity. Accordingly, the intrinsic complexity of the cell–matrix–cell interactions of the NVU has made analyzing gene function in cell culture, tissue explants, and even animal models difficult and the inability to study gene function in the BBB in vivo has been a critical hurdle to advancing BBB research. Zebrafish has emerged as a premier vertebrate organism to model and analyze complex cellular interactions in vivo and genetic mechanisms of embryonic development. To this end, we provide a technical overview of the procedures that can be used in Zebrafish to analyze BBB integrity with a focus on the cerebrovasculature of adult fish where the BBB is now defined. The techniques that are used to measure the functional integrity, the cell biology, and the ultrastructure of the BBB include permeability assays, fluorescent imaging of reporter genes, and electron microscopy, respectively. Each can be applied to the functional analysis of mutant fish in ways that characterize the molecular sequelae to pathological insults that compromise BBB integrity. Due to the highly conserved nature of both the genetics and cell biology of zebrafish when compared with higher vertebrates, drug discovery techniques can be used in zebrafish models to complement drug development studies in other model systems.