Visualization of vesicle transport along and between distinct pathways in neurites of living cells

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Abstract
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 ob- served consistently, pointing to a functional relevance of vesicle cotransport. Microsc. Res. Tech. 63: 159 -167, 2004. © 2004 Wiley-Liss, Inc.
Funding Information
  • Austrian Federal Ministry of Science and Transport (GZ 200.025/3-Pr/4/1998, GZ 200.027/3-Pr/2a/1998)
  • Austrian Research Funds (P12803-MED, P15053)