ODORANT-RECEPTOR INTERACTIONS IN OLFACTORY NEURONS

这项研究计划中描述的实验的长期目标 是为了研究化学感觉的综合生理活性 在细胞、亚细胞和分子水平上的粘膜。特别的 兴趣是分泌细胞及其产物的作用，包括 糖基化分泌型糖偶联物及其信息素 转运蛋白vomomodrin与受体周围事件相关 化学传感转导。实现这一目标的具体目标是： 答：研究调节人类健康的外在因素和内在因素。 鼻侧腺体发育过程中腭裂调节蛋白的表达 使用分子生物学技术，包括原位杂交和 Northern印迹分析，并将其与对照进行比较。 推测的气味转运体气味结合蛋白；B.测定 N-乙酰氨基葡萄糖分泌物糖基化的亚细胞位置 并研究了它们在电子致密的和非电子的 -粘液的透明粘液结构域和分子外域 用标记的凝集素探针检测与嗅觉纤毛相关的糖萼 用金微球；C.研究了金微球的微观结构 嗅觉粘膜和犁鼻粘膜的粘膜感觉间隔 使用凝集素探针和抗体的激光共聚焦扫描显微镜 用荧光染料标记；以及D.检查发育和 气味调控的I相表达和II相生物转化 酶，定位于支持细胞和分泌细胞以及 粘膜感受器，包括细胞色素P450NMA/b、谷胱甘肽 S转移酶和γ-谷氨酰转肽酶的免疫细胞化学研究 和分子生物学技术。成功执行了 拟议的实验将产生基本的基本科学见解 进入与周受体相关的分子调控机制 化学感觉转导研究进展及临床研究进展 与嗅觉障碍和水和 粘液性鼻漏。

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.