Kaul Aamna K, Bateman Joseph M
Wolfson Centre for Age-Related Diseases, King's College London, UK.
J Vis Exp. 2009 Dec 15(34):1617. doi: 10.3791/1617.
The Drosophila eye is a powerful model system for studying areas such as neurogenesis, signal transduction and neurodegeneration. Many of the discoveries made using this system have taken advantage of the spatiotemporal nature of photoreceptor differentiation in the developing eye imaginal disc. To use this system it is first necessary for the researcher to learn to identify and dissect the eye disc. We describe a novel RFP reporter to aid in the identification of the eye disc and the visualization of specific cell types in the developing eye. We detail a methodology for dissection of the eye imaginal disc from third instar larvae and describe how the eye-RFP reporter can aid in this dissection. This eye-RFP reporter is only expressed in the eye and can be visualized using fluorescence microscopy either in live tissue or after fixation without the need for signal amplification. We also show how this reporter can be used to identify specific cells types within the eye disc. This protocol and the use of the eye-RFP reporter will aid researchers using the Drosophila eye to address fundamentally important biological questions.
果蝇眼睛是研究神经发生、信号转导和神经退行性变等领域的强大模型系统。利用该系统取得的许多发现都得益于发育中的眼成虫盘光感受器分化的时空特性。要使用这个系统,研究人员首先需要学会识别和解剖眼盘。我们描述了一种新型红色荧光蛋白(RFP)报告基因,以帮助识别眼盘并可视化发育中眼睛的特定细胞类型。我们详细介绍了从三龄幼虫解剖眼成虫盘的方法,并描述了眼-RFP报告基因如何有助于这种解剖。这种眼-RFP报告基因仅在眼睛中表达,可在活组织中或固定后使用荧光显微镜进行可视化,无需信号放大。我们还展示了该报告基因如何用于识别眼盘内的特定细胞类型。本方案以及眼-RFP报告基因的使用将有助于研究人员利用果蝇眼睛解决具有根本重要性的生物学问题。