Effects of dynein inhibitor on the number of motor proteins transporting synaptic cargos.

Synaptic cargo transport by kinesin and dynein in hippocampal neurons was investigated by noninvasively measuring the transport force based on nonequilibrium statistical mechanics. Although direct physical measurements such as force measurement using optical tweezers are difficult in an intracellular environment, the noninvasive estimations enabled enumerating force-producing units (FPUs) carrying a cargo comprising the motor proteins generating force. The number of FPUs served as a barometer for stable and long-distance transport by multiple motors, which was then used to quantify the extent of damage to axonal transport by dynarrestin, a dynein inhibitor. We found that dynarrestin decreased the FPU for retrograde transport more than for anterograde transport. This result indicates the applicability of the noninvasive force measurements. In the future, these measurements may be used to quantify damage to axonal transport resulting from neuronal diseases, including Alzheimer's, Parkinson's, and Huntington's diseases.

Physical parameters describing neuronal cargo transport by kinesin UNC-104.

In this review, we focus on the kinesin-3 family molecular motor protein UNC-104 and its regulatory protein ARL-8. UNC-104, originally identified in Caenorhabditis elegans (C. elegans), has a primary role transporting synaptic vesicle precursors (SVPs). Although in vitro single-molecule experiments have been performed to primarily investigate the kinesin motor domain, these have not addressed the in vivo reality of the existence of regulatory proteins, such as ARL-8, that control kinesin attachment to/detachment from cargo vesicles, which is essential to the overall transport efficiency of cargo vesicles. To quantitatively understand the role of the regulatory protein, we review the in vivo physical parameters of UNC-104-mediated SVP transport, including force, velocity, run length and run time, derived from wild-type and arl-8-deletion mutant C. elegans. Our future aim is to facilitate the construction of a consensus physical model to connect SVP transport with pathologies related to deficient synapse construction caused by the deficient UNC-104 regulation. We hope that the physical parameters of SVP transport summarized in this review become a useful guide for the development of such model.