Methods for Quantitative Analysis of Axonal Cargo Transport

Abstract

Neurons rely on complex axonal transport mechanisms that mediate the intracellular dynamics of proteins, vesicles, and mitochondria along their high polarized structure. The fast improvement of live imaging techniques of fluorescent cargos allowed the identification of the diverse motion properties of different transported molecules. These properties arise as the result of molecular interactions between many players involved in axonal transport. Motor proteins, microtubule tracks, cargo association, and even axonal viscosity contribute to the proper axonal dynamics of different cargos. The unique properties in each cargo determine their distribution and location that is relevant to ensure neuronal cell activity and survival. This chapter provides a computational-based method for the generation of cargo trajectories and the identification of different motion regimes while cargo moves along axons. Then, the procedure to extract relevant parameters from active, diffusive, and confined motion is provided. These properties will allow a better comprehension of the nature and characteristics of cargo motion in living cells, therefore contributing to understanding the consequences of transport defects that arise during diseases of the nervous system.

Keywords: Anterograde; Axonal transport; Diffusive; Pauses; Retrograde; Reversions; Run lengths; Segmental velocities.