Fellows, A. D. | bioRxiv Science

bioRxiv2023

Dynein and dynactin move long-range but are delivered separately to the axon tip

Axonal transport is essential for neuronal survival. This is driven by microtubule motors including dynein, which transports cargo from the axon tip back to the cell body. This function requires its cofactor dynactin and regulators LIS1 and NDEL1. Due to difficulties imaging dynein at a single-molecule level, it is unclear how this motor and its regulators coordinate transport along the length of the axon. Here we use neuron-inducible human stem-celllines (NGN2-OPTi-OX) to endogenously tag dynein components and visualise them at a near-single molecule regime. In the retrograde direction, we find that dynein and dynactin can move the entire length of the axon (>500 m) in one go. Furthermore, LIS1 and NDEL1 also undergo longdistance movement, despite being mainly implicated with initiation of dynein transport. Intriguingly, in the anterograde direction, dynein/LIS1 move faster than dynactin/NDEL1 consistent with transport on different cargos. Therefore, neurons ensure efficient transport by holding dynein/dynactin on cargos over long distances, but keeping them separate until required.

cell biology↗

bioRxiv2022

Bimodal regulation of axonal transport by the GDNF-RET signalling axis in healthy and diseased motor neurons

Deficits in axonal transport are one of the earliest pathological outcomes in several models of amyotrophic lateral sclerosis (ALS), including SOD1G93A mice. Evidence suggests that rescuing these deficits prevents disease progression, stops denervation, and extends survival. Kinase inhibitors have been previously identified as transport enhancers, and are being investigated as potential therapies for ALS. For example, inhibitors of p38 mitogen-activated protein kinase and insulin growth factor receptor 1 have been shown to rescue axonal transport deficits in vivo in symptomatic SOD1G93A mice. In this work, we investigated the impact of RET, the tyrosine kinase receptor for glial cell-line-derived neurotrophic factor (GDNF), as a modifier of axonal transport. We identified fundamental interplay between RET signalling and axonal transport in both wild type and SOD1G93A motor neurons in vitro. We demonstrated that blockade of RET signalling using pharmacological inhibitors and genetic knockdown enhances signalling endosome transport in wild type motor neurons and uncovered a divergence in the response of primary motor neurons to GDNF compared with cell lines. Finally, we demonstrated that inhibition of the GDNF-RET signalling axis rescues in vivo transport deficits in early symptomatic SOD1G93A mice, promoting RET as a potential therapeutic target in the treatment of ALS.

neuroscience↗