Bingham Stephanie M, Toussaint Gesulla, Chandrasekhar Anand
Division of Biological Sciences, and Molecular Biology Program, Room 205 Lefevre Hall, University of Missouri, Columbia, MO 65211, USA.
J Neurosci Methods. 2005 Nov 30;149(1):42-9. doi: 10.1016/j.jneumeth.2005.05.002. Epub 2005 Jun 20.
The zebrafish embryo is an excellent system for studying dynamic processes such as cell migration during vertebrate development. Dynamic analysis of neuronal migration in the zebrafish hindbrain has been hampered by morphogenetic movements in vivo, and by the impermeability of embryos. We have applied a recently reported technique of embryo explant culture to the analysis of neuronal development and migration in the zebrafish hindbrain. We show that hindbrain explants prepared at the somitogenesis stage undergo normal morphogenesis for at least 14 h in culture. Importantly, several aspects of hindbrain development such as patterning, neurogenesis, axon guidance, and neuronal migration are largely unaffected, inspite of increased cell death in explanted tissue. These results suggest that hindbrain explant culture can be employed effectively in zebrafish to analyze neuronal migration and other dynamic processes using pharmacological and imaging techniques.
斑马鱼胚胎是研究脊椎动物发育过程中细胞迁移等动态过程的优秀系统。体内形态发生运动以及胚胎的不渗透性阻碍了对斑马鱼后脑神经元迁移的动态分析。我们将最近报道的胚胎外植体培养技术应用于斑马鱼后脑神经元发育和迁移的分析。我们发现,在体节形成阶段制备的后脑外植体在培养中至少14小时能经历正常的形态发生。重要的是,尽管外植组织中的细胞死亡增加,但后脑发育的几个方面,如模式形成(patterning)、神经发生、轴突导向和神经元迁移在很大程度上不受影响。这些结果表明,后脑外植体培养可有效地用于斑马鱼,以利用药理学和成像技术分析神经元迁移及其他动态过程。