Dynamic imaging of axonal transport in living XXXX cells in vitro.

@entity2136 @entity2479 @entity5611 @entity1781 @entity3812 @entity35 @entity204
PURPOSE: Disruption of the axonal transport of neurotrophic factors plays a critical role in the apoptosis of @entity2479 cells (RGCs) in @entity1781 . Live-cell imaging in vitro was used in this study, to visualize and evaluate the axonal transport of @entity2136 ( @entity2136 ) in both living and axon-damaged RGCs. METHODS: @entity35 RGCs were purified by a two-step immunopanning method. Cultivated RGCs were transfected with a plasmid encoding @entity2136 tagged with green fluorescent protein (GFP), and the dynamics of @entity2136 in the axons and dendrites were analyzed by time-lapse imaging. Changes in the axonal transport of @entity2136 were examined after treatment with 1 mM @entity3812 , and @entity204 after treatment was evaluated with @entity5611 homodimer-1. RESULTS: The expression of @entity2136 showed a vesicular pattern in the axons and dendrites of cultivated RGCs. Time-lapse imaging revealed that the mean velocity of @entity2136 in the axons (0.86    0.37  m/s; maximum = 2.03  m/s) was significantly greater (P < 0.0001) than that in the dendrites (mean = 0.49    0.19  m/s). @entity3812 significantly inhibited the axonal transport of @entity2136 -GFP at 2 and 3 hours after treatment (P = 0.003 and 0.0002, respectively) without affecting cell viability; however, @entity204 was detected 24 hours after treatment. CONCLUSIONS: Live-cell imaging revealed the dynamics of the axonal transport of @entity2136 in living RGCs, which clearly differed from the movements in dendrites. Furthermore, it was possible to confirm the disruption of axonal transport in @entity3812 -treated RGCs before cell death.
