Rosenbaum Corinna, Böttner Martina, Wedel Thilo, Metzger Marco
Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg, Würzburg, Germany.
Department of Anatomy, University of Kiel, Kiel, Germany.
Methods Mol Biol. 2018;1723:361-369. doi: 10.1007/978-1-4939-7558-7_20.
The enteric nervous system (ENS) is the division of the autonomic nervous system that innervates the gastrointestinal (GI) tract and controls central intestinal functions such as peristalsis and fluid movement. Enteric nerve cell bodies (neurons and glia) are predominantly organized in ganglionated networks that are present along the entire length of the GI tract in multiple tissue layers. Most cell bodies are organized in the myenteric plexus allocated between the longitudinal and the circular muscle layers or in the submucosal plexus between muscle tissue and mucosa. The site-specific characteristics of these enteric nerve cells have traditionally been analyzed via imaging techniques. Laser-capture microdissection (LCM) offers the prospect of site-specifically analyzing the gene expression profiles of these different subpopulations. This protocol addresses critical aspects of handling intestinal tissue for ENS dissection, such as the optimal quick-staining procedure, suitable laser settings, and limits of tissue material required to successfully dissect and analyze tissue layers for gene expression.
肠神经系统（ENS）是自主神经系统的一个分支，它支配胃肠道（GI）并控制诸如蠕动和液体流动等肠道核心功能。肠神经细胞体（神经元和神经胶质细胞）主要组织成神经节网络，这些网络存在于胃肠道全长的多个组织层中。大多数细胞体组织在位于纵肌层和环肌层之间的肌间神经丛中，或在肌肉组织和黏膜之间的黏膜下神经丛中。传统上，这些肠神经细胞的位点特异性特征是通过成像技术进行分析的。激光捕获显微切割（LCM）为位点特异性分析这些不同亚群的基因表达谱提供了可能。本方案阐述了处理用于肠神经系统解剖的肠道组织的关键方面，例如最佳快速染色程序、合适的激光设置，以及成功解剖和分析组织层以进行基因表达所需的组织材料限度。
