The Identification of Green Fluorescence Protein Tagged Kinesin Mutations in Caenorhabditis Elegans Through Fluorescence Microscopy
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Information is passed between neurons and muscles through the release of neurotransmitters. These neurotransmitters are stored in synaptic vesicles in synapses. But before they can dock at a synapse, a motor transport known as kinesin moves the synaptic vesicles along microtubules to their final destination in the cell. This process is known as vesicle transport.
The purpose of these experiments were to gain a better understanding of the movement of vesicles along microtubules and the importance of this in proper nerve transmission. This was accomplished by analyzing the phenotypes of three different strains of Caenorhabditis elegans using microscopy and a fluorescence microscope. The organisms contained fluorescence tags on the membranes of their synaptic vesicles to make it easier to see how the mutations affected the vesicles locations.
Wild type C. elegans should be actively moving in an s-shape pattern under a microscope. Under a fluorescence microscope, the tags should appear at the nerve ring and along the nerve cords. In the C. elegans with a fusion defect, its movement would appear impaired and the fluorescence tags would be more concentrated at the nerve ring. Lastly, a C. elegans with a transport defect would also have impaired movement, but the fluorescence tag would be dispersed in patches along the length of the entire worm.
Observation of phenotypic appearance was taken using a dissecting microscope at 50x magnification and a