ODORANT-RECEPTOR INTERACTIONS IN OLFACTORY NEURONS

本研究提案中描述的实验的长期目标 是研究化学感受器的综合生理活动， 在细胞、亚细胞和分子水平上对粘膜的影响。 特别 感兴趣的是分泌细胞及其产物的作用，包括 糖基化分泌糖复合物和推定的信息素 犁骨调节蛋白转运蛋白，在与 化学感受性转导 实现这一目标的具体目标是： a：研究调节 犁骨调节蛋白在侧鼻腺发育中的表达 使用分子生物学技术，包括原位杂交， 北方杂交分析，并与 推定的气味剂转运蛋白气味剂结合蛋白; B.确定 N-乙酰葡糖胺糖基化亚细胞位点 糖缀合物，并检查它们在电子致密和 - 粘液的透明粘液样结构域和 用凝集素探针标记的与嗅觉纤毛相关的糖萼 具有金微球; c.为了研究 嗅觉和犁鼻粘膜的粘膜感觉区室， 使用凝集素探针和抗体的共聚焦激光扫描显微术 用荧光染料标记;和d.研究发展和 I期和II期生物转化的气味调节表达 酶，定位于支持细胞和分泌细胞以及 粘膜感觉区室，包括细胞色素P450 NMa/B、谷胱甘肽 免疫细胞化学法检测S-转移酶和γ-谷氨酰转肽酶 和分子生物学技术。 成功执行 提出的实验将产生基本的基础科学见解 与受体周围相关的分子调节机制 化学感受转导中的事件和临床见解， 与嗅觉丧失和水样和 粘液性溢液。

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.