Author:
Chowdary Praveen D.,McGuire Allister F.,Lee Yujeong,Che Daphne L.,Hanson Lindsey,Osakada Yasuko,Ooi Chinchun,Xie Chong,Wang Shan,Cui Bianxiao
Abstract
AbstractNoninvasive control of axonal cargos in live neurons is a challenging prospect that can enable novel research on the mechanisms of axonal cargo transport, cargo-mediated signaling and axonal traffic jams in neurons. However, conventional techniques for force manipulation such as optical traps are limited to a few micron-sized cargos and are not applicable to the small axonal cargos in live neurons. Here, we present a new methodology that permits the external control of axonal endosome transport via tailored magnetic forces. By culturing neurons in a microfluidic device made up of microfabricated magnetic arrays, we can exert 3 – 48 pN forces on retrograde axonal endosomes carrying fluorescent magnetic nanoparticles, 100 – 260 nm in size. The magnetic force counters the forces exerted by molecular motors driving the endosomes and results in a wide range of perturbations on endosome transport in axons. These perturbations, captured by oblique illumination fluorescence imaging, reveal new insights on the collective function of dyneins and the nature of paused and stationary states during retrograde endosome transport in axons. Most notably, we demonstrate controllable capture and release of retrograde endosomes in axons by toggling the external magnetic field. This technical advance has great potential to elucidate the spatiotemporal origins of long-distance endosome signaling pathways as well as the ramifications of axonal traffic jams in neurons.
Publisher
Cold Spring Harbor Laboratory
Reference42 articles.
1. Axonal Transport: Components, Mechanisms, and Specificity
2. Stenoien, D. L. & Brady, S. T. Axonal transport. Basic neurochemistry: Molecular, cellular and medical aspects, Sixth edition, 565–587, (1999).
3. Molecular motors in axonal transport
4. Kinesin molecular motors: Transport pathways, receptors, and human disease
5. Retrograde axonal transport: pathways to cell death?
Cited by
7 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献