Visualization of vesicle transport along and between distinct pathways in neurites of living cells.
Trafficking of secretory vesicles along neurites of PC12 cells was visualized by 2D and 3D real-time imaging using fluorescence microscopy. Vesicle motion along distinct pathways was directly seen. From an overlay of individual pathways, the underlying cytoskeletal filament could be imaged at a subwavelength resolution. Continuous vesicle transport was interrupted by periods of diffusive motion with concomitant pathway changes. Statistical analysis shows that such interruptions were distributed stochastically along the filament, indicating a limited processivity of motor proteins also in a cellular context. Periods of diffusive motion facilitated the interaction with actively transported vesicles. Frequent associations and dissociations of vesicles have been observed consistently, pointing to a functional relevance of vesicle cotransport.
Schütz
Gerhard J
GJ
Institute for Biophysics, University of Linz, A-4040 Linz, Austria. gerhard.schuetz@jku.at
Axmann
Markus
M
Freudenthaler
Susanne
S
Schindler
Hansgeorg
H
Kandror
Kostya
K
Roder
John C
JC
Jeromin
Andreas
A
D000818:Animals; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D016501:Neurites / Q000378:metabolism / Q000648:ultrastructure*; D016716:PC12 Cells; D021381:Protein Transport; D051381:Rats
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins