Using Live-Cell Imaging to Measure the Effects of Pathological Proteins on Axonal Transport in Primary Hippocampal Neurons

Neurons rely on bidirectional transport of cargo along the axon to maintain functional synapsis and neural connectivity. Deficits in transport are thought to be critical contributors to the pathogenesis of several neurodegenerative diseases. We aim to identify mechanisms by which disease-associated modifications to tau protein disrupt axonal transporting.

We established that multiple pathological forms of tau protein disrupt normal axonal transport in mammalian neurons. This effect depends on changes to phosphorylation-based signaling pathways that regulate axonal transport. These disruptions represent a potential mechanism of tau-induced neurotoxicity in disease.

Modified forms of specific proteins can disrupt normal fast axonal transport across several neurodegenerative diseases. However, we don't fully understand the underlying mechanisms of these effects. This provides a reproducible and simple protocol to identify how expression of disease associated proteins affect fast axonal transport in mammalian neurons.

This protocol provides a reproducible assay for axonal transport in mammalian primary neurons, that is easily modifiable to include various cargo proteins along with the expression of proteins of interest with specific pathological modifications. It also allows for the targeted knockdown of other pathway components to test particular mechanistic details.