Journal
CYTOSKELETON
Volume 75, Issue 1, Pages 22-41Publisher
WILEY
DOI: 10.1002/cm.21411
Keywords
axon; axonal transport; kinetics; kymograph; neurofilament
Categories
Funding
- NIH [R01 NS038526, S10 OD010383, P30 NS045758]
- NSF [IOS 1656784, IOS 1656785]
- Neuroscience Research Institute at The Ohio State University
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We have used kymograph analysis combined with edge detection and an automated computational algorithm to analyze the axonal transport kinetics of neurofilament polymers in cultured neurons at 30 ms temporal resolution. We generated 301 kymographs from 136 movies and analyzed 726 filaments ranging from 0.6 to 42 mu m in length, representing approximate to 37,000 distinct moving and pausing events. We found that the movement is even more intermittent than previously reported and that the filaments undergo frequent, often transient, reversals which suggest that they can engage simultaneously with both anterograde and retrograde motors. Average anterograde and retrograde bout velocities (0.9 and 1.2 mu ms(-1), respectively) were faster than previously reported, with maximum sustained bout velocities of up to 6.6 and 7.8 mu ms(-1), respectively. Average run lengths (approximate to 1.1 mu m) and run times (approximate to 1.4 s) were in the range reported for molecular motor processivity in vitro, suggesting that the runs could represent the individual processive bouts of the neurofilament motors. Notably, we found no decrease in run velocity, run length or run time with increasing filament length, which suggests that either the drag on the moving filaments is negligible or that longer filaments recruit more motors.
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