ANALYSIS OF ODORANT RECEPTOR GENES

描述(摘自申请者摘要)：该项目旨在揭示 嗅觉系统功能和发展的基本原则，通过 新近发现的气味受体DOR家族的详细分析 基因。该提案涉及受体功能的基本问题，服用 果蝇作为实验系统的优势。嗅觉 是一种古老的感觉形式，嗅觉功能和许多原理 无脊椎动物和脊椎动物之间的组织结构保存得很好。此外， 了解昆虫的嗅觉可能有助于控制昆虫媒介 通过嗅觉找到人类宿主或配偶的人类疾病 暗示。 该提案旨在首先对DOR基因的所有成员进行识别和分类 家庭，包括高度不同的成员。影响DOR的现有突变 人们将寻找基因。 本课程将讨论成熟嗅觉系统中受体的分布。 在两个方面。首先，最近分离出了一种针对DOR蛋白的抗体 允许对嗅觉受体的定位进行高分辨率分析 系统。结果表明，DOR22A.2蛋白存在于空洞中 嗅觉感受器，正如预期的那样，定位于树突上的气味感受器， 将通过免疫电子显微镜和抗体来确认和推广 对抗额外的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。 提出了一种受体表达的发育分析。发展中的 将分析所有DOR基因的图谱，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 嗅觉感受器神经元的轴突将得到确认和延伸。 对选定的DOR基因的遗传分析被用来检验这一假说 它们编码气味感受器。DOR基因功能将通过 过度表达和功能丧失突变。这些实验可能 确定DOR蛋白的配基。他们还可以提供功能性证据。 关于每个神经元表达的受体的数量，以及 受体的发育作用。

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.