Ames Laboratory, US Department of Energy, Ames, Iowa 50011, USA.
Nat Commun. 2012;3:1030. doi: 10.1038/ncomms2037.
Direct visualization of axonal transport in live neurons is essential for our understanding of the neuronal functions and the working mechanisms of microtubule-based motor proteins. Here we use the high-speed single particle orientation and rotational tracking technique to directly visualize the rotational dynamics of cargos in both active directional transport and pausing stages of axonal transport, with a temporal resolution of 2 ms. Both long and short pauses are imaged, and the correlations between the pause duration, the rotational behaviour of the cargo at the pause, and the moving direction after the pause are established. Furthermore, the rotational dynamics leading to switching tracks are visualized in detail. These first-time observations of cargo's rotational dynamics provide new insights on how kinesin and dynein motors take the cargo through the alternating stages of active directional transport and pause.
直接可视化活神经元中的轴突运输对于我们理解神经元功能和基于微管的运动蛋白的工作机制至关重要。在这里，我们使用高速单颗粒取向和旋转跟踪技术，以 2 毫秒的时间分辨率直接可视化轴突运输的主动定向运输和暂停阶段中货物的旋转动力学。我们对长暂停和短暂停都进行了成像，并建立了暂停持续时间、暂停时货物的旋转行为以及暂停后移动方向之间的相关性。此外，还详细观察了导致切换轨迹的旋转动力学。这些对货物旋转动力学的首次观察为驱动蛋白和动力蛋白如何使货物通过主动定向运输和暂停的交替阶段提供了新的见解。
