Molecular Mechanisms of Specific Anosmia

描述（由申请人提供）：本项目将通过研究雄烯酮和其他配体的特异性嗅觉和致敏性，重点研究哺乳动物嗅觉的分子机制。雄甾酮气味受体(s)将使用筛选系统匹配受体与其同源配体。通过对受体转运蛋白（rtp）的结构-功能分析，将进一步加强这一系统。将通过细胞表面免疫染色和荧光素酶测定来评估rtp的缺失和嵌合序列，以确定关键区域。rtp和气味受体之间的相互作用域将使用分裂-泛素酵母双杂交系统进行研究。候选雄烯酮气味受体的多态性将通过心理物理分析人群的基因分型来评估其与嗅觉表型的相关性。雄甾酮嗅觉型和嗅觉型小鼠模型将用于相似的SNP基因型分析，以及mRNA和蛋白表达分析。体内敏化和异源细胞实验将揭示配体暴露如何促进雄烯酮的敏化。

DESCRIPTION (provided by applicant): The project outlined in this application will focus on the molecular mechanism underlying mammalian odor perception through the investigations of specific anosmia and sensitization of androstenone and other ligands. Androstenone odorant receptor(s) will be identified using a screening system matching receptors to their cognate ligands. Such system will be strengthened by a structure-function analysis of the receptor- transporting proteins (RTPs). Deletion and chimeric series of RTPs will be assessed in cell-surface immunostaining and luciferase assay to identify key regions. Domains of interaction between RTPs and odorant receptors will be investigated using the split-ubiquitin yeast two-hybrid system. The polymorphisms in candidate androstenone odorant receptor(s) will be evaluated for correlation with the anosmic phenotype by genotyping a psychophysically-analyzed population. Mouse models of androstenone-anosmic and osmic strains will be used for similar SNP genotype analysis, as well as mRNA and protein expression analyses. In vivo sensitization and experiments in heterologous cell will shed lights how ligand exposure facilitates androstenone sensitization.