Dong Audrey, Kulkarni Vineet Vinay, Maday Sandra
Department of Neuroscience, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
Methods Mol Biol. 2019;1880:243-256. doi: 10.1007/978-1-4939-8873-0_16.
Autophagy is an essential degradative pathway that maintains neuronal homeostasis and prevents axon degeneration. However, the mechanisms of autophagy in neurons are only beginning to be understood. To address this fundamental gap in knowledge, we have established several key methodologies for live-cell imaging and quantitative analysis of autophagy in primary hippocampal neurons. Using these methods, we have defined compartment-specific dynamics of autophagy in real-time under basal versus stress conditions. For example, we have characterized autophagosome biogenesis in the distal axon and subsequent retrograde transport to the soma for degradation. Autophagosomes are also generated locally within the soma. In contrast to the axon, the majority of autophagosomes in dendrites are stationary, while some exhibit bidirectional movement. These studies establish an initial road map for autophagosome dynamics in each compartment of the neuron and set the stage for a more detailed understanding of neuronal autophagy in stress and disease.
自噬是一种重要的降解途径，可维持神经元内环境稳定并防止轴突退化。然而，自噬在神经元中的机制才刚刚开始被了解。为了填补这一知识上的根本空白，我们建立了几种用于原代海马神经元自噬的活细胞成像和定量分析的关键方法。使用这些方法，我们已经确定了在基础状态与应激条件下自噬在不同区域的实时动态变化。例如，我们已经描绘了远端轴突中自噬体的生物发生过程以及随后向胞体的逆行运输以便进行降解。自噬体也在胞体局部产生。与轴突不同，树突中的大多数自噬体是静止的，而有些则表现出双向运动。这些研究为神经元每个区域的自噬体动态变化建立了初步路线图，并为更详细地了解应激和疾病状态下的神经元自噬奠定了基础。
