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Single-particle fluorescence tracking combined with TrackMate assay reveals highly heterogeneous and discontinuous lysosomal transport in freely orientated axons
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  • Yongyang Liu,
  • Yaxin Lu,
  • Zhiyong Tang,
  • Yuheng Cao,
  • Dehua Huang,
  • Feng Wu,
  • Yejun Zhang,
  • Chunyan Li,
  • Guangcun Chen,
  • Qiangbin Wang
Yongyang Liu
University of Science and Technology of China
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Yaxin Lu
University of Science and Technology of China
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Zhiyong Tang
University of Science and Technology of China
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Yuheng Cao
ShanghaiTech University
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Dehua Huang
Suzhou Institute of Nano-tech and Nano-Bionics Chinese Academy of Sciences
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Feng Wu
Suzhou Institute of Nano-tech and Nano-Bionics Chinese Academy of Sciences
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Yejun Zhang
Suzhou Institute of Nano-tech and Nano-Bionics Chinese Academy of Sciences
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Chunyan Li
Suzhou Institute of Nano-tech and Nano-bionics
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Guangcun Chen
Suzhou Institute of Nano-tech and Nano-bionics
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Qiangbin Wang
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Abstract

Axonal transport plays a significant role in the establishment of neuronal polarity, axon growth, and synapse formation during neuronal development. The axon of a naturally growing neuron is a highly complex and multifurcated structure with a large number of bends and branches. Nowadays, the study of dynamic axonal transport in morphologically complex neurons is greatly limited by the technological barrier. Here, a sparse gene transfection strategy was developed to locate fluorescent mCherry in the lysosome of primary neurons, thus enabling us to track the lysosome-based axonal transport with a single-particle resolution. Thereby, several axonal transport models were observed, including forward or backward transport model, stop-and-go model, repeated back-and-forth transport model, and cross-branch transport model. Then, the accurate single-particle velocity quantification by TrackMate revealed a highly heterogeneous and discontinuous transportation process of lysosome-based axonal transport in freely orientated axons. And, multiple physical factors, such as the axonal structure and the size of particles, were disclosed to affect the velocity of particle transporting in freely orientated axons. The combined single-particle fluorescence tracking and TrackMate assay can be served as a facile tool for evaluating axonal transport in neuronal development and axonal transport-related diseases.
04 Jan 2022Submitted to Biotechnology Journal
05 Jan 2022Submission Checks Completed
05 Jan 2022Assigned to Editor
14 Jan 2022Reviewer(s) Assigned
12 Feb 2022Editorial Decision: Revise Major
11 Jun 20221st Revision Received
13 Jun 2022Submission Checks Completed
13 Jun 2022Assigned to Editor
18 Jun 2022Reviewer(s) Assigned
22 Jun 2022Editorial Decision: Accept