Functional and genetic analysis of choroid plexus development in zebrafish
Functional and genetic analysis of choroid plexus development in zebrafish
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DOI：
10.3389/fnins.2014.00364
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发表时间：
2014-11-10
影响因子：
4.3
通讯作者：
Taylor, Michael R.
中科院分区：
文献类型：
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作者：
Henson, Hannah E.;Parupalli, Chaithanyarani;Taylor, Michael R.
脉络丛是位于脑室的上皮结构，是血-脑脊液屏障的主要组成部分。脉络丛产生脑脊髓液并调节脑脊髓液的成分。脉络丛功能异常与神经退行性疾病、脉络丛上皮肿瘤形成和脑积水有关。在这项研究中，我们使用斑马鱼（Danio rerio）作为模型系统，以了解脉络丛发育的遗传成分。我们产生了增强子捕获线，Et（cp：EGFP）（sj 2），在脉络丛上皮中表达增强型绿色荧光蛋白（EGFP）。利用免疫组织化学和荧光示踪，我们证明了斑马鱼脉络丛具有脑屏障的性质，如紧密连接和转运活性。因此，我们建立了斑马鱼作为功能相关的模型，研究脉络丛的发展。使用无偏的方法，我们进行了正向遗传解剖脉络丛，以确定其形成和功能所必需的基因。使用Et（cp：EGFP）（sj 2），我们分离了10个具有脉络丛异常的隐性突变株系，基于GFP强度、上皮定位和总体脉络丛形态将其分为五类。我们还将两个突变株系的突变分别定位到第4和21号染色体上。本研究中产生的突变体可用于阐明脉络丛发育，功能和/或维护所必需的特定基因和信号通路，并将为这些基因突变如何导致疾病提供重要见解。
The choroid plexus, an epithelial-based structure localized in the brain ventricle, is the major component of the blood-cerebrospinal fluid barrier. The choroid plexus produces the cerebrospinal fluid and regulates the components of the cerebrospinal fluid. Abnormal choroid plexus function is associated with neurodegenerative diseases, tumor formation in the choroid plexus epithelium, and hydrocephaly. In this study, we used zebrafish(Danio rerio) as a model system to understand the genetic components of choroid plexus development. We generated an enhancer trap line, Et(cp: EGFP)(sj2), that expresses enhanced green fluorescent protein(EGFP) in the choroid plexus epithelium. Using immunohistochemistry and fluorescent tracers, we demonstrated that the zebrafish choroid plexus possesses brain barrier properties such as tight junctions and transporter activity. Thus, we have established zebrafish as a functionally relevant model to study choroid plexus development. Using an unbiased approach, we performed a forward genetic dissection of the choroid plexus to identify genes essential for its formation and function. Using Et(cp: EGFP)(sj2), we isolated 10 recessive mutant lines with choroid plexus abnormalities,which were grouped into five classes based on GFP intensity, epithelial localization, and overall choroid plexus morphology. We also mapped the mutation for two mutant lines to chromosomes 4 and 21, respectively. The mutants generated in this study can be used to elucidate specific genes and signaling pathways essential for choroid plexus development, function, and/or maintenance and will provide important insights into how these genetic mutations contribute to disease.