Comparative analysis of axonal transport markers in primary mammalian neurons

Axonal transport is important for neuronal development and the maintenance of effective neuronal function in mature cells. Observing the active transport of organelles and vesicles along the axons of living neurons has emerged as a valuable tool for probing the health of the neuron, and assessing changes associated with stress and neurodegenerative disease. Transport relies on two families of motor proteins: kinesins and dynein. Using these motors, a diverse set of cargos are transported toward the axon tip, the cell body, or anywhere in between. Of particular interest are organelles and cargos associated with disease and the changes in motility that these cargos undergo during pathogenesis.Here, we describe the factors that should be considered when studying different cargos, and the imaging parameters associated with optimal tracking of various organelles and proteins. Ultimately, the ideal cargo to investigate depends on the question being asked and the limitations of individual microscopes available for imaging.