%AFenn, J. [Department of Neuroscience and Medical Scientist Training Program Ohio State University Columbus Ohio 43210]%AJohnson, Christopher [Quantitative Biology Institute and Department of Physics and Astronomy Ohio University Athens Ohio 45701]%APeng, Juan [Center for Biostatistics and Department of Biomedical Informatics Ohio State University Columbus Ohio 43210]%AJung, Peter [Quantitative Biology Institute and Department of Physics and Astronomy Ohio University Athens Ohio 45701]%ABrown, Anthony [Department of Neuroscience and Medical Scientist Training Program Ohio State University Columbus Ohio 43210]%BJournal Name: Cytoskeleton; Journal Volume: 75; Journal Issue: 1; Related Information: CHORUS Timestamp: 2023-09-06 09:23:24 %D2017%IWiley Blackwell (John Wiley & Sons) %JJournal Name: Cytoskeleton; Journal Volume: 75; Journal Issue: 1; Related Information: CHORUS Timestamp: 2023-09-06 09:23:24 %K %MOSTI ID: 10046914 %PMedium: X %TKymograph analysis with high temporal resolution reveals new features of neurofilament transport kinetics %XAbstract

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 µm in length, representing ∼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 µm s−1, respectively) were faster than previously reported, with maximum sustained bout velocities of up to 6.6 and 7.8 µm s−1, respectively. Average run lengths (∼1.1 µm) and run times (∼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.

%0Journal Article