Molecular Analysis of Axonal Transport Dynamics upon Modulation of Microtubule Acetylation.

Axonal transport is used by neurons to distribute mRNAs, proteins, and organelles to their peripheral compartments in order to sustain their structural and functional integrity. Cargoes are transported along the microtubule (MT) network whose post-translational modifications influence transport dynamics. Here, we describe methods to modulate MT acetylation and record its impact on axonal transport in cultured mouse cortical projection neurons as well as in motoneurons of Drosophila melanogaster third-instar larvae. Specifically, we provide a step-by step procedure to reduce the level of MT acetylation and to record and analyze the transport of dye-labeled organelles in projection neuron axons cultured in microfluidic chambers. In addition, we describe the method to record and analyze GFP-tagged mitochondria transport along the motoneuron axons of transgenic Drosophila melanogaster third-instar larvae.