This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The C. elegans neuromuscular junction is a model synapse for the study of synapse development and function. We are combining genetics with physiological and morphological tools to characterize the C. elegans neuromuscular junction. Over the past year we have refined a protocol which preserves fine structure at synapses, such as microtubules, synaptic vesicles and filamentous elements, for analysis under the electron beam. Specifically, we embed intact worms in resin after high-pressure freeze immobilization and freeze substitution in acetone with tannic acid and, secondarily, osmium. Currently, we are conducting detailed morphometric analysis and would like to extend our analysis to 3-D reconstructions derived from high-voltage electron microscopy.
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