ODORANT-RECEPTOR INTERACTIONS IN OLFACTORY NEURONS

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The long-term goal of the experiments described in this research proposal is to investigate the integrated physiological activity of chemosensory mucosae at the cellular, subcellular and molecular levels. Of particular interest is the role of secretory cells and their products, including glycosylated secretory glycoconjugates and the putative pheromone transporter vomeromodulin, in perireceptor events associated with chemosensory transduction. The specific aims to achieve this goal are: a: to study the extrinsic and intrinsic factors that regulate the developmental expression of vomeromodulin by the lateral nasal glands using molecular biological techniques including in situ hybridization and Northern blot analysis, and to compare the regulation with that of putative odorant transporter odorant binding protein; b. to determine the subcellular site of N-acetylglucosamine glycosylation of secretory glycoconjugates and to examine their distribution in electron-dense and - lucent mucoid domains of mucus and in the molecular ectodomains of the glycocalyx associated with olfactory cilia using lectinoprobes labeled with gold microspheres; c. to investigate the microstructure of the mucosensory compartments of the olfactory and vomeronasal mucosae with confocal laser scanning microscopy using lectinoprobes and antibodies labeled with fluorochromes; and d. to examine the developmental and odorant-regulated expression of Phase I and Phase II biotransformation enzymes, localized in sustentacular and secretory cells as well as mucosensory compartments, including cytochrome P450 NMa/b, glutathione S-transferases and gamma-glutamyl transpeptidase using immunocytochemical and molecular biological techniques. Successful execution of the proposed experiments will yield fundamental basic scientific insights into the molecular regulatory mechanisms associated with perireceptor events in chemosensory transduction and clinical insights into chemosensory transport disorders associated with anosmia and watery and mucoid rhinorrhea.