用于在临床前阿尔茨海默病模型中研究神经元细胞间信号相互作用的双克隆系统方案。
Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer's disease model.
作者信息
Singh Aditi, Deshpande Prajakta, Sarkar Ankita, Yeates Catherine, Chimata Anuradha Venkatakrishnan, Kango-Singh Madhuri, Singh Amit
机构信息
Department of Biology, University of Dayton, Dayton, OH 45469, USA.
Department of Biology, University of Dayton, Dayton, OH 45469, USA; Premedical Program, University of Dayton, Dayton, OH, USA; Integrative Science and Engineering (ISE), University of Dayton, Dayton, OH, USA; Center for Genomic Advocacy (TCGA), Indiana State University, Terre Haute, IN, USA.
出版信息
STAR Protoc. 2025 Aug 8;6(3):103993. doi: 10.1016/j.xpro.2025.103993.
This protocol outlines a two-clone genetic mosaic approach in Drosophila to study cell signaling using the FLP/FRT (Flippase/Flippase recombination target) and Gal4/UAS (upstream activating sequence)/Gal80 systems. We generated a stable transgenic line expressing human Aβ42, and then we crossed it with a GFP-marked FRT line to produce mosaic clones via heat-shock-induced recombination. Then we dissected, stained, and imaged the eye discs to visualize wild-type and mutant clones. Image analysis with Fiji/ImageJ and statistical quantification in GraphPad Prism enable the study of cell competition and intercellular signaling mechanism(s) in human diseases. For complete details on the use and execution of this protocol, please refer to Yeates et al. and Tare et al..
本方案概述了在果蝇中使用FLP/FRT（翻转酶/翻转酶重组靶点）和Gal4/UAS（上游激活序列）/Gal80系统研究细胞信号传导的双克隆基因镶嵌方法。我们构建了一个表达人Aβ42的稳定转基因品系，然后将其与一个绿色荧光蛋白标记的FRT品系杂交，通过热休克诱导重组产生镶嵌克隆。然后我们解剖、染色并对眼盘进行成像，以观察野生型和突变克隆。使用Fiji/ImageJ进行图像分析以及在GraphPad Prism中进行统计定量，有助于研究人类疾病中的细胞竞争和细胞间信号传导机制。有关本方案使用和实施的完整详细信息，请参考Yeates等人以及Tare等人的文献。
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