Dumas Laura, Durand Jason, Loulier Karine
Institute for Neuroscience of Montpellier (INM), University of Montpellier, INSERM, Montpellier, France.
Department of Medical Neuroscience, Brain Repair Centre, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada.
Methods Mol Biol. 2025;2886:47-63. doi: 10.1007/978-1-0716-4310-5_3.
Multicolor MAGIC Markers strategies are useful lineage tracing tools to study brain development at a multicellular scale. In this chapter, we describe an in utero electroporation method to simultaneously label multiple neighboring progenitors and their respective progeny using these multicolor reporters. In utero electroporation enables the introduction of any gene of interest into embryonic neural progenitors lining the brain ventricles through a simple pipeline consisting of a micro-injection followed by the application of electrical pulses. Successful in utero electroporation requires a concise yet complete understanding of each step of the surgical protocol, spanning from the preoperative preparation to the postoperative care, as well as the MAGIC Markers tool outlined in this study. Besides a detailed protocol, we present non-integrative and integrative approaches to demonstrate the range of cell and lineage tracking possibilities of multicolored progenitors and their descent over time.
多色MAGIC标记策略是在多细胞水平研究大脑发育的有用谱系追踪工具。在本章中，我们描述了一种子宫内电穿孔方法，使用这些多色报告基因同时标记多个相邻的祖细胞及其各自的子代。子宫内电穿孔能够通过一个简单流程，即将显微注射与电脉冲应用相结合，将任何感兴趣的基因导入衬于脑室的胚胎神经祖细胞中。成功的子宫内电穿孔需要对手术方案的每一步骤有精确且全面的理解，涵盖术前准备到术后护理，以及本研究中概述的MAGIC标记工具。除了详细的方案，我们还介绍了非整合和整合方法，以展示多色祖细胞及其后代随时间推移的细胞和谱系追踪可能性范围。
