ANALYSIS OF ODORANT RECEPTOR GENES

气味受体基因分析

• 批准号: 6489590
• 负责人: John R Carlson
• 金额: $ 37.17万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6489590
• 关键词: Drosophilidae; biological signal transduction; brain mapping; chemoreceptors; developmental genetics; electrophysiology; functional /structural genomics; gene mutation; growth /development; immunocytochemistry; ligands; molecular genetics; morphometry; neurophysiology; odors; olfactions; olfactory stimulus; olfactory threshold; protein localization; protein structure function; receptor binding; receptor expression; sensory discrimination; sensory signal detection; serial analysis of gene expression

DESCRIPTION (From the Applicant's Abstract): This project is designed to reveal basic principles of olfactory system function and development, through a detailed analysis of the recently discovered DOR family of odorant receptor genes. The proposal addresses fundamental issues of receptor function, taking advantage of the strengths of Drosophila as an experimental system. Olfaction is an ancient sensory modality, and many principles of olfactory function and organization are well conserved among invertebrates and vertebrates. Moreover, understanding of insect olfaction may be useful in controlling insect vectors of human disease that find their human hosts or their mates through olfactory cues. The proposal aims first to identify and classify all members of the DOR gene family, including highly divergent members. Extant mutations affecting DOR genes will be sought. The distribution of receptors in the mature olfactory system will be addressed in two ways. First, the recent isolation of an antibody against a DOR protein allows a high-resolution analysis of receptor localization in the olfactory system. Results showing that the DOR22A.2 protein is present in the cavities of olfactory sensilla, as expected of an odorant receptor localized to dendrites, will be confirmed and extended by immunoelectron microscopy and with antibodies against additional DOR proteins. Second, the number of receptors per neuron will be investigated by double-label in situ hybridization. A developmental analysis of receptor expression is proposed. The developmental profiles of all DOR genes will be analyzed, with a view to determining how many genes are expressed early in development, as is DOR22A.2. Preliminary immunohistochemical results indicating that the DOR22A.2 receptor is present on the axons of olfactory receptor neurons will be confirmed and extended. A genetic analysis of selected DOR genes is proposed to test the hypothesis that they encode odorant receptors. DOR gene function will be altered through overexpression and by loss-of-function mutations. These experiments may identify a ligand for a DOR protein. They may also provide functional evidence regarding the number of receptors expressed per neurons, and the possibility of developmental roles for receptors.

描述（来自申请人的摘要）：该项目旨在揭示 嗅觉系统的功能和发展的基本原则，通过一个 对最近发现的气味受体DOR家族的详细分析 基因.该提案涉及受体功能的基本问题， 果蝇作为实验系统的优势。嗅觉 嗅觉是一种古老的感觉方式，嗅觉功能的许多原理， 组织在无脊椎动物和脊椎动物中保存良好。此外，委员会认为， 对昆虫嗅觉的了解可能有助于控制昆虫媒介 通过嗅觉找到人类宿主或配偶 线索 该提案旨在首先识别和分类DOR基因的所有成员 家庭，包括高度分歧的成员。影响DOR的现存突变 基因将被寻找。 受体的分布在成熟的嗅觉系统将解决 从两个方面。首先，最近分离出了针对DOR蛋白的抗体， 允许高分辨率分析嗅觉中的受体定位 系统结果显示DOR22A.2蛋白存在于大肠杆菌的腔中， 嗅觉感受器，如定位于树突的气味受体所预期的， 将通过免疫电子显微镜和抗体进行确认和扩展 对抗其他DOR蛋白第二，每个神经元的受体数量 将通过双标记原位杂交进行研究。 提出了受体表达的发育分析。发育 将分析所有DOR基因的概况，以确定有多少 基因在发育早期表达，DOR22A.2也是如此。初步 免疫组织化学结果表明DOR22A.2受体存在于 嗅觉受体神经元的轴突将被确认和延伸。 选择DOR基因的遗传分析，提出了测试的假设 它们编码气味受体。DOR基因的功能将被改变， 过度表达和功能丧失突变。这些实验可能 鉴定DOR蛋白配体。他们还可以提供功能证据 关于每个神经元表达的受体数量，以及 受体的发育作用。