Neurons are large, complex cells that use a microtubule motor protein-based system for the fast transport of proteins and organelle cargo throughout long, complex projections. In output projections known as axons, cytoplasmic dynein transports cargo in the retrograde direction towards the cell body. Dynein requires the activating complex dynactin, including the subunit DCTN1 (p150glued). DCTN1 is post-translationally modified with the lipid palmitate in a process known as palmitoylation at cysteines 617 and 1255, likely by ZDHHC12. Additionally, DCTN1 is preferentially palmitoylated in the nervous system. In neurons, when wild type (wt) or palmitoylation resistant (C617,1255A; CCAA) are ectopically expressed, there may be more wtDCTN1-GFP in distal axons than DCTN1-CCAA-GFP. The results of this study provide a greater understanding of the regulation of transport in neurons with implications for synaptic function and neurodegeneration.