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      Characterization of the movement of the kinesin motor KIF1A in living cultured neurons.

      The Journal of Biological Chemistry
      Animals, Biological Transport, Brain, embryology, COS Cells, Cell Line, Cells, Cultured, Green Fluorescent Proteins, Hippocampus, cytology, Humans, Image Processing, Computer-Assisted, Kinesin, chemistry, metabolism, Luminescent Proteins, Microscopy, Fluorescence, Microtubules, Movement, Nerve Tissue Proteins, Neurons, Precipitin Tests, Protein Structure, Tertiary, Rats, Recombinant Fusion Proteins, Time Factors, Transfection

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          Abstract

          KIF1A is a kinesin motor known to transport synaptic vesicle precursors in neuronal axons, but little is known about whether KIF1A mediates fast and processive axonal transport in vivo. By monitoring movements of EGFP-labeled KIF1A in living cultured hippocampal neurons, we determined the characteristics of KIF1A movements. KIF1A particles moved anterogradely along the neurites with an average velocity of 1.0 microm/s. The movements of KIF1A were highly processive, with an average duration of persistent anterograde movement of 11 s. Some KIF1A particles (17%) exhibited retrograde movements of 0.72 microm/s, although overall particle movement was in the anterograde direction. The anterograde movement of KIF1A, however, did not lead to a detectable accumulation of KIF1A in the periphery of neurons, suggesting that there are mechanisms inhibiting the peripheral accumulation of KIF1A. These results suggest that KIF1A mediates neuronal transport at a high velocity and processivity in vivo.

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