Axonal actin in action: Imaging actin dynamics in neurons

Methods in Cell Biology
Kelsey LadtSubhojit Roy

Abstract

Actin is a highly conserved, key cytoskeletal protein involved in numerous structural and functional roles. In neurons, actin has been intensively investigated in axon terminals-growth cones-and dendritic spines, but details about actin structure and dynamics in axon shafts have remained obscure for decades. A major barrier in the field has been imaging actin. Actin exists as soluble monomers (G-actin) as well as actin filaments (F-actin), and labeling actin with conventional fluorescent probes like GFP/RFP typically leads to a diffuse haze that makes it difficult to discern kinetic behaviors. In a recent publication, we used F-actin selective probes to visualize actin dynamics in axons, resolving striking actin behaviors that have not been described before. However, using these probes to visualize actin dynamics is challenging as they can cause bundling of actin filaments; thus, experimental parameters need to be strictly optimized. Here we describe some practical methodological details related to using these probes for visualizing F-actin dynamics in axons.

Citations

Jul 29, 2015·The Journal of Cell Biology·Archan GangulySubhojit Roy
Jun 14, 2019·Physical Biology·Nilaj Chakrabarty, Peter Jung
Nov 8, 2018·The Journal of Cell Biology·Nilaj ChakrabartySubhojit Roy
Nov 27, 2018·F1000Research·Shalini Menon, Stephanie Gupton
Apr 6, 2021·Frontiers in Oncology·Xin JiangYanhong Zhou

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