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The Role of Microtubule End Binding Proteins in Axon and
Synapse Development in C. elegans Microtubule associated protein localization and interactions at the subcellular level can be studied with the help of fluorescent proteins. Cell-specific promoters target expression of the fluorescent protein-tagged transgenes to particular cell types in the developing animal. In this study, we used GFP fused to a synaptic vesicle protein (synaptobrevin-GFP) to examine synapses in two end binding protein deletion mutants, klp-7(tm2143) and ebp-2(gk737). The synaptobrevin-GFP marker (snb-1::GFP) was crossed into each mutant, and snb-1::GFP patterns were then compared to the expression of snb-1::GFP in wild-type animals. The subcellular localization of an ebp-2::GFP fusion protein was also studied by constructing animals that expressed ebp-2::GFP and unc-10::RFP or ebp-2::GFP and snb-1::RFP. unc-10 encodes the C. elegans RIM protein, a known active zone protein. ebp-2::GFP is expressed in a punctate pattern, but did not co-localize directly with unc-10::RFP or snb-1::GFP, indicating that it is most likely present in the periactive zone surrounding the active zone of the synapse. Support provided by: National Science Foundation-Research at Undergraduate Institutions Grant to Prof. Baran |
