Direct visualization of the movement of the monomeric axonal transport motor UNC-104 along neuronal processes in living Caenorhabditis elegans
| Autor: | Ingrid Brust-Mascher, Jonathan M. Scholey, H. Mimi Zhou |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Nervous system
Time Factors Recombinant Fusion Proteins Axon terminus Green Fluorescent Proteins Gene Expression Nerve Tissue Proteins Biology Motor Activity Axonal Transport Green fluorescent protein Motor protein Microtubule medicine Animals ARTICLE Caenorhabditis elegans Caenorhabditis elegans Proteins Neurons General Neuroscience Molecular Motor Proteins Gene Transfer Techniques Kymography Dendrites Axons Cell biology Luminescent Proteins medicine.anatomical_structure Phenotype nervous system Microscopy Fluorescence Organ Specificity Axoplasmic transport Kinesin Synaptic vesicle transport |
| Zdroj: | The Journal of neuroscience : the official journal of the Society for Neuroscience. 21(11) |
| ISSN: | 1529-2401 |
| Popis: | The formation and function of axons depends on the microtubule-based transport of cellular components from their sites of synthesis in the neuronal cell body to their sites of utilization at the axon terminus. To directly visualize this axonal transport in a living organism, we constructed transgenic lines ofCaenorhabditis elegansthat express green fluorescent protein fused to the monomeric synaptic vesicle transport motor, UNC-104. This UNC-104:: GFP construct rescued the Unc-104 mutant phenotype and was expressed throughout the nervous system. Using time-lapse confocal fluorescence microscopy, we were able to visualize fluorescent motor proteins moving in both directions along neuronal processes, some of which were identified definitely as axons and others as dendrites. Using kymograph analysis, we followed the movement of >900 particles. Most of them moved in one direction, but not necessarily at the same velocity. Ten percent of the observed particles reversed direction of movement during the period of observation, and 10% exhibited periods of movement interspersed with pauses. During episodes of persistent movement, particles moved at an average velocity of 1.02 μm/sec, which is close to thein vitrovelocity of microtubule gliding driven by purified monomeric kinesin at high motor density. To our knowledge, this is the first direct visualization and analysis of the movement of specifically labeled microtubule motor proteins along axonsin vivo. |
| Databáze: | OpenAIRE |
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