Walther Straub Institute of Pharmacology and Toxicology, Faculty of Medicine, LMU Munich, Munich, Germany.
Department of Human Biology, Technical University Munich, Freising-Weihenstephan, Germany.
Methods Mol Biol. 2022;2510:145-156. doi: 10.1007/978-1-0716-2384-8_8.
P2X7 receptors play an important role in cytokine release and immune cell regulation. Their upregulation has been described in inflammatory and degenerative processes and P2X7 blockade or deletion has been shown to reduce tissue damage and severity of symptoms in animal models of inflammatory bowel disease (IBD). Several studies have found that P2X7 receptors are present on enteric neurons and glia and it was proposed that they mediate neuronal death during IBD. However, the cell type-specific localization of P2X7 receptors has been a matter of debate, since some antibodies have been found to be unspecific. Here we describe the preparation of whole-mount myenteric plexus from the colon of BAC transgenic P2X7-EGFP reporter mice and subsequent immunofluorescence staining of P2X7 receptors together with cell type-specific marker proteins.
P2X7 受体在细胞因子释放和免疫细胞调节中发挥重要作用。它们在炎症和退行性过程中的上调已被描述，并且已经证明 P2X7 阻断或缺失可以减少炎症性肠病 (IBD) 动物模型中的组织损伤和症状严重程度。几项研究发现 P2X7 受体存在于肠神经元和神经胶质细胞中，有人提出它们在 IBD 期间介导神经元死亡。然而，P2X7 受体的细胞类型特异性定位一直存在争议，因为一些抗体被发现是特异性的。在这里，我们描述了从 BAC 转基因 P2X7-EGFP 报告小鼠的结肠制备全层肌间神经丛，并随后对 P2X7 受体进行免疫荧光染色，同时对细胞类型特异性标记蛋白进行染色。
