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. Work was started in the past year to apply the glycomics profiling platform being developed at the Resource to the characterization of GAGs in the C. elegans model organism.
The aim of this project is to develop the technology necessary to perform GAG glycomics experiments. The technology will be developed using C. elegans because of the demonstrated importance of this organism as a model for studying proteoglycan functions (9). CS/DS is known to be essential for C. elegans reproductive maturation (vulval development), and disruption of genes essential to its attachment to proteins is operative in the phenotype of a series of reproductive mutants (squashed vulva, or sqv mutants). Heparan sulfate proteoglycans are known to be necessary for the developmental architecture in C. elegans, and disruption of HS processing enzymes for the correct positioning of axons along the ventral midline (10, 11). Given the established roles of CS and HS in C. elegans development, a detailed investigation of the expression of GAGs is needed. The technology will then be applied to larval stages of C. elegans. Initial activity in this project has focused on optimizing procedures for isolation of GAGs from the nematode in a form appropriate for subsequent capillary electrophoretic disaccharide and electrospray MS analyses.
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