Visualizing Vesicle-Bound Kinesins in Cultured Hippocampal Neurons.

Andrew Montgomery, Alex Garbouchian, Marvin Bentley

Methods in molecular biology (Clifton, N.J.) 2022

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Eukaryotic cells use microtubule-based vesicle transport to exchange molecules between compartments. Kinesin family members mediate all microtubule plus end-directed vesicle transport. Of the 45 kinesins expressed in humans, some 20 mediate microtubule plus-end directed vesicle transport. Here we describe a technique to visualize vesicle-bound kinesins in cultured hippocampal neurons. The method involves the expression of the vesicle-binding tail domain while minimizing the cytoplasmic pool. Using this approach drastically improves vesicle labeling compared to full-length kinesins. This tool is useful for systematically comparing the localization of different kinesins in the same cell type and for identifying cargo proteins that reside in vesicles moved by a specific kinesin family member. While we describe the assay in cultured hippocampal neurons, we expect it to be easily transferable to other eukaryotic cell types. © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Andrew Montgomery, Alex Garbouchian, Marvin Bentley. Visualizing Vesicle-Bound Kinesins in Cultured Hippocampal Neurons. Methods in molecular biology (Clifton, N.J.). 2022;2431:239-247