Figure 3.
Dynein moves long-range along the axon in a stable complex. (A) Example kymograph of retrograde dynein (Halo-DYNC1H1) movement in 21–23 DPI neurons treated with either JFX 554 or JFX 650, see also Fig. S3. (B) The run lengths of retrograde dynein particles in 21–23 DPI neurons. (C) The speed of retrograde dynein particles in 21–23 DPI neurons (162 tracks, 21 videos, N = 6). (D) Schematic of the experimental setup to explore dynein movement in the axon. (1) JFX dyes were added to the axon tip at T = 0 min. This labels both diffusive and motile dynein. (2) Motile dynein moves along the axon. Dynein either forms a stable interaction with cargo and moves the entire length of the axon or undergoes short-range movements and then dissociates from the complex. (3) Our imaging window at the proximal end of the axon. If dynein is stably bound, we expected to see the first fluorescent spot within 5 min. On the other hand, if dynein is exchanged on cargo, it should take longer. (E) The amount of time until the first processive fluorescent particle was detected in dynein (Halo-DYNC1H1) 21–23 DPI neurons (8 videos, N = 8). Boxplots shows median, first, and third quartiles. Upper/lower whiskers extend to 1.5× the interquartile range.

Dynein moves long-range along the axon in a stable complex. (A) Example kymograph of retrograde dynein (Halo-DYNC1H1) movement in 21–23 DPI neurons treated with either JFX 554 or JFX 650, see also Fig. S3. (B) The run lengths of retrograde dynein particles in 21–23 DPI neurons. (C) The speed of retrograde dynein particles in 21–23 DPI neurons (162 tracks, 21 videos, N = 6). (D) Schematic of the experimental setup to explore dynein movement in the axon. (1) JFX dyes were added to the axon tip at T = 0 min. This labels both diffusive and motile dynein. (2) Motile dynein moves along the axon. Dynein either forms a stable interaction with cargo and moves the entire length of the axon or undergoes short-range movements and then dissociates from the complex. (3) Our imaging window at the proximal end of the axon. If dynein is stably bound, we expected to see the first fluorescent spot within 5 min. On the other hand, if dynein is exchanged on cargo, it should take longer. (E) The amount of time until the first processive fluorescent particle was detected in dynein (Halo-DYNC1H1) 21–23 DPI neurons (8 videos, N = 8). Boxplots shows median, first, and third quartiles. Upper/lower whiskers extend to 1.5× the interquartile range.

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