Visualization and manipulation of the platelet and megakaryocyte cytoskeleton.
Visualization and manipulation of the platelet and megakaryocyte cytoskeleton.
复制标题
DOI：
10.1007/978-1-61779-307-3_9
复制
发表时间：
2012
影响因子：
--
通讯作者：
Jonathan N Thon;J. Italiano
中科院分区：
文献类型：
--
作者：
Jonathan N Thon;J. Italiano
免疫荧光（IF）显微镜和现代分子生物学方法的细胞骨架操作的应用程序的驱动下，在过去的5年里取得了相当大的进展，我们了解巨核细胞发育和血小板生物发生的分子机制。这些研究已经可视化了内有丝分裂纺锤体动力学，表征了分界膜系统的成熟，描绘了活巨核细胞中细胞器运输和微管组装的机制，描述了体内血小板产生的过程，并揭示了促进前血小板产生和血小板释放的因素和驱动机制。在这里，我们描述的方法（1）培养巨核细胞从小鼠胎肝，（2）操纵微管蛋白和肌动蛋白细胞骨架的血小板和培养的巨核细胞，（3）检查这些活细胞显微镜和固定细胞免疫荧光显微镜。
Driven by the application of immunofluorescence (IF) microscopy and modern molecular biology approaches to cytoskeletal manipulation, the last 5 years have yielded considerable progress to our understanding of the molecular mechanisms governing megakaryocyte development and platelet biogenesis. Such studies have visualized endomitotic spindle dynamics, characterized the maturation of the demarcation membrane system, delineated the mechanics of organelle transport and microtubule assembly in living megakaryocytes, described the process of platelet production in vivo, and revealed factors contributing to and the mechanisms driving proplatelet production and platelet release. Here, we describe methods to (1) culture megakaryocytes from murine fetal livers, (2) manipulate the tubulin and actin cytoskeleton of both platelets and cultured megakaryocytes, and (3) examine these by live-cell microscopy and fixed-cell immunofluorescence microscopy.