Super-Resolution Microscopy Reveals the Native Ultrastructure of the Erythrocyte Cytoskeleton
Super-Resolution Microscopy Reveals the Native Ultrastructure of the Erythrocyte Cytoskeleton
复制标题
超分辨率显微镜揭示红细胞细胞骨架的天然超微结构
DOI：
10.1016/j.celrep.2017.12.107
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发表时间：
2018-01-30
期刊：
影响因子：
8.8
通讯作者：
Xu, Ke
中科院分区：
文献类型：
--
作者：
Pan, Leiting;Yan, Rui;Xu, Ke
红细胞骨架是后生动物细胞膜下细胞骨架的教科书原型。虽然早期的实验表明，肌动蛋白为基础的连接复合物的三角形网络连接类似200 nm长的血影蛋白四聚体，后来的研究表明，小得多的连接到连接的距离在25-60 nm的范围内。通过超分辨率显微镜，我们解决了天然超微结构的细胞骨架的膜保存的红细胞的N和C末端的b-血影蛋白，F-肌动蛋白，蛋白4.1，原调节蛋白，和内收蛋白。这使我们能够确定一个类似的80 nm的交界处到交界处的距离，长度与放松血影蛋白四聚体和理论的基础上血影蛋白丰度一致。通过双色数据，我们进一步表明，细胞骨架网络往往包含纳米级的空隙，细胞膜保持完整，肌动蛋白丝和帽蛋白定位于一个子集，但不是所有的，连接复合物。总之，我们的研究结果要求重新评估的结构和功能的膜下细胞骨架。
The erythrocyte cytoskeleton is a textbook prototype for the submembrane cytoskeleton of metazoan cells. While early experiments suggest a triangular network of actin-based junctional complexes connected by similar to 200-nm-long spectrin tetramers, later studies indicate much smaller junction-to-junction distances in the range of 25-60 nm. Through super-resolution microscopy, we resolve the native ultrastructure of the cytoskeleton of membrane-preserved erythrocytes for the N and C termini of b-spectrin, F-actin, protein 4.1, tropomodulin, and adducin. This allows us to determine an similar to 80-nm junction-to-junction distance, a length consistent with relaxed spectrin tetramers and theories based on spectrin abundance. Through two-color data, we further show that the cytoskeleton meshwork often contains nanoscale voids where the cell membrane remains intact and that actin filaments and capping proteins localize to a subset of, but not all, junctional complexes. Together, our results call for a reassessment of the structure and function of the submembrane cytoskeleton.