A P2X1-eYFP knock-in mouse was generated to study receptor expression and mobility in smooth muscle and blood cells. eYFP was added to the C-terminus of the P2X1R and replaced the native P2X1R. Fluorescence corresponding to P2X1-eYFPR was detected in urinary bladder smooth muscle, platelets and megakaryocytes. ATP-evoked currents from wild type and P2X1-eYFP isolated urinary bladder smooth muscle cells had the same peak current amplitude and time-course showing that the eYFP addition had no obvious effect on properties. Fluorescence recovery after photobleaching (FRAP) in bladder smooth muscle cells demonstrated that surface P2X1Rs are mobile and their movement is reduced following cholesterol depletion. Compared to the platelet and megakaryocyte, P2X1-eYFP fluorescence was negligible in red blood cells and the majority of smaller marrow cells. The spatial pattern of P2X1-eYFP fluorescence in the megakaryocyte along with FRAP assessment of mobility suggested that P2X1Rs are expressed extensively throughout the membrane invagination system of this cell type. The current study highlights that the spatiotemporal properties of P2X1R expression can be monitored in real time in smooth muscle cells and megakaryocytes/platelets using the eYFP knock-in mouse model.

中文翻译：

---

开发P2X1-eYFP受体敲入小鼠以实时跟踪受体。

产生了一只P2X1-eYFP敲入小鼠来研究受体在平滑肌和血细胞中的表达和迁移。eYFP被添加到P2X1R的C末端，并取代了天然P2X1R。在膀胱平滑肌，血小板和巨核细胞中检测到对应于P2X1-eYFPR的荧光。来自野生型和P2X1-eYFP分离的膀胱平滑肌细胞的ATP诱发电流具有相同的峰值电流幅度和时间过程，表明添加eYFP对性能没有明显影响。膀胱平滑肌细胞光漂白后的荧光恢复（FRAP）表明，表面P2X1Rs可移动，胆固醇耗尽后其运动减少。与血小板和巨核细胞相比，P2X1-eYFP荧光在红细胞和大多数较小的骨髓细胞中可忽略不计。巨核细胞中P2X1-eYFP荧光的空间模式以及对迁移率的FRAP评估表明，P2X1Rs在该细胞类型的整个膜侵入系统中广泛表达。当前的研究强调，可以使用eYFP敲入小鼠模型实时监测平滑肌细胞和巨核细胞/血小板中P2X1R表达的时空特性。