Visualization of clathrin-mediated endocytosis during Semaphorin-guided axonal growth
Visualization of clathrin-mediated endocytosis during Semaphorin-guided axonal growth
复制标题
信号蛋白引导的轴突生长过程中网格蛋白介导的内吞作用的可视化
DOI：
10.1007/978-1-4939-6448-2_21
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发表时间：
2017
影响因子：
--
通讯作者：
Hiroyuki Kamiguchi
中科院分区：
文献类型：
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作者：
Rurika Itofusa;Takuro Tojima;Hiroyuki Kamiguchi
Semaphorin3A（Sema3A）在神经元网络发育期间指导轴突生长。越来越多的证据表明Sema3A诱导的生长锥塌陷和排斥涉及生长锥中的内吞膜运输。现在可以使用全内反射荧光显微镜（TIRFM）来可视化活细胞中的内吞过程，TIRFM是一种用于在质膜上成像动态亚细胞事件的强大工具。在这一章中，我们描述了一种方法，TIRFM观察和分析网格蛋白介导的内吞作用在鸡背根神经节神经元的生长锥，接收细胞外浓度梯度的Sema3A在培养基中。
Semaphorin3A (Sema3A) guides axonal growth during neuronal network development. Accumulating evidence indicates that Sema3A-induced growth cone collapse and repulsion involve endocytic membrane trafficking in the growth cone. It is now possible to visualize endocytic processes in living cells using total internal reflection fluorescence microscopy (TIRFM), a powerful tool for imaging dynamic subcellular events at the plasma membrane. In this chapter, we describe a method for TIRFM observation and analysis of clathrin-mediated endocytosis in growth cones of chicken dorsal root ganglion neurons that receive an extracellular concentration gradient of Sema3A in a culture medium.