Rentzsch Fabian, Renfer Eduard, Technau Ulrich
Sars International Centre for Marine Molecular Biology, University of Bergen, Bergen, Norway.
Department of Biological Sciences, University of Bergen, Bergen, Norway.
Methods Mol Biol. 2020;2047:45-57. doi: 10.1007/978-1-4939-9732-9_3.
Neurons often display complex morphologies with long and fine processes that can be difficult to visualize, in particular in living animals. Transgenic reporter lines in which fluorescent proteins are expressed in defined populations of neurons are important tools that can overcome these difficulties. By using membrane-attached fluorescent proteins, such reporter transgenes can identify the complete outline of subsets of neurons or they can highlight the subcellular localization of fusion proteins, for example at pre- or postsynaptic sites. The relative stability of fluorescent proteins furthermore allows the tracing of the progeny of cells over time and can therefore provide information about potential roles of the gene whose regulatory elements are controlling the expression of the fluorescent protein. Here we describe the generation of transgenic reporter lines in the sea anemone Nematostella vectensis, a cnidarian model organism for studying the evolution of developmental processes. We also provide an overview of existing transgenic Nematostella lines that have been used to study conserved and derived aspects of nervous system development.
神经元通常呈现出具有长而精细突起的复杂形态，这可能难以可视化，尤其是在活体动物中。荧光蛋白在特定神经元群体中表达的转基因报告系是克服这些困难的重要工具。通过使用膜附着荧光蛋白，此类报告转基因可以识别神经元亚群的完整轮廓，或者突出融合蛋白的亚细胞定位，例如在突触前或突触后位点。此外，荧光蛋白的相对稳定性允许随时间追踪细胞后代，因此可以提供有关其调控元件控制荧光蛋白表达的基因潜在作用的信息。在这里，我们描述了在海葵星状海葵（Nematostella vectensis）中生成转基因报告系的过程，海葵是一种用于研究发育过程进化的刺胞动物模型生物。我们还概述了现有的用于研究神经系统发育保守和衍生方面的转基因星状海葵系。
