Knutson Katilyn, Strege Peter R, Li Joyce, Leiter Andrew B, Farrugia Gianrico, Beyder Arthur
Enteric Neuroscience Program, Division of Gastroenterology & Hepatology,Department of Physiology & Biomedical Engineering, Mayo Clinic.
Division of Gastroenterology, Department of Medicine, University of Massachusetts Medical School.
J Vis Exp. 2018 Sep 26(139):58112. doi: 10.3791/58112.
Enterochromaffin (EC) cells in the gastrointestinal (GI) epithelium constitute the largest subpopulation of enteroendocrine cells. As specialized sensory cells, EC cells sense luminal stimuli and convert them into serotonin (5-hyroxytryptamine, 5-HT) release events. However, the electrophysiology of these cells is poorly understood because they are difficult to culture and to identify. The method presented in this paper outlines primary EC cell cultures optimized for single cell electrophysiology. This protocol utilizes a transgenic cyan fluorescent protein (CFP) reporter to identify mouse EC cells in mixed primary cultures, advancing the approach to obtaining high-quality recordings of whole cell electrophysiology in voltage- and current-clamp modes.
胃肠道（GI）上皮中的肠嗜铬（EC）细胞是肠内分泌细胞中最大的亚群。作为特殊的感觉细胞，EC细胞感知腔内刺激并将其转化为血清素（5-羟色胺，5-HT）释放事件。然而，由于这些细胞难以培养和识别，它们的电生理学尚不清楚。本文介绍的方法概述了针对单细胞电生理学优化的原代EC细胞培养。该方案利用转基因青色荧光蛋白（CFP）报告基因来识别混合原代培养物中的小鼠EC细胞，改进了在电压钳和电流钳模式下获得全细胞电生理学高质量记录的方法。
