MOLECULAR BIOLOGY OF OLFACTORY RECEPTOR GENES

嗅觉受体基因的分子生物学

• 批准号: 6175418
• 负责人: Andrew J Chess
• 金额: $ 44.25万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6175418
• 关键词: alleles; animal genetic material tag; chemoreceptors; gene rearrangement; genetic regulatory element; genetically modified animals; laboratory mouse; neurogenetics; olfactions; receptor expression; tissue /cell culture; vomeronasal systems

Understanding the development and function of the brain requires the elucidation of mechanisms underlying the generation of an extremely large number of neurons with unique identities. Moreover, the multitude of distinct neurons ultimately must connect to each other in specific patterns. The olfactory system provides an excellent model system for investigating these questions because the 1,000 olfactory receptor genes serve as molecular markers for functionally distinct subsets of neurons. Our preliminary studies indicate that a hierarchy of controls operates on the family of olfactory receptor genes, such that a given olfactory neuron expresses only one of the thousand possible genes. One mechanism involved in regulating olfactory receptor gene expression is allelic inactivation (only one of two alleles is expressed). Based on these studies, the aims of this proposal are to elucidate mechanisms underlying olfactory receptor gene regulation and allelic inactivation: 1. To determine if DNA rearrangement plays a role in the choice of which olfactory receptor is expressed. 2. To define the cis-acting DNA elements involved in regulating olfactory receptor genes. 3. To further characterize allelic inactivation and to identify its underlying molecular mechanisms. The potential role of rearrangement will be addressed in molecular studies of purified olfactory neurons expressing a given receptor and in cell lines derived from the olfactory neuroepithelium. Once cells which have chosen a given receptor are isolated, the genomic DNA will be analyzed to look for DNA rearrangement. Independent of the question of rearrangement, we wish to understand elements of control. We will use yeast artificial chromosomes (YACs) to map the loci encoding olfactory receptors. Cis acting DNA elements involved in receptor gene choice will be defined in transgenic mouse experiments. Experiments to characterize allelic inactivation will involve analyses of transcription and DNA replication in cell lines and transgenic mice. These studies will elucidate mechanisms through which gene regulation leads to the remarkable generation of neuronal diversity in the brain, and will be of value in understanding disorders of brain function. Moreover, mammalian olfactory neurons regenerate throughout the life of the organism. Thus, understanding the gene expression mechanisms used by these neurons is of particular relevance to understanding neurodevelopmental disorders.

了解大脑的发展和功能需要 阐明产生极大的机制 具有独特身份的神经元数。 而且，众多 不同的神经元最终必须以特定的方式相互连接 模式。 嗅觉系统为 研究这些问题是因为1,000个嗅觉受体基因 充当神经元功能上不同子集的分子标记。 我们的初步研究表明，控制的层次结构 嗅觉受体基因的家族，使给定的嗅觉神经元 仅表示千种可能的基因中的一个。 一种机制 在调节嗅觉受体基因表达中是等位基因失活 （表示两个等位基因中的一个）。 基于这些研究，目的 该提议是为了阐明嗅觉受体的基础机制 基因调节和等位基因失活： 1。确定DNA重排是否在选择中起作用 表达嗅觉受体。 2。定义与调节嗅觉有关的顺式作用DNA元素 受体基因。 3。进一步描述等位基因失活并确定其 潜在的分子机制。 重排的潜在作用将在分子研究中解决 表达给定受体和细胞系的纯化的嗅觉神经元 源自嗅觉神经上皮。 曾经选择的细胞 分离给定受体，将分析基因组DNA以寻找 DNA重排。 独立于重排的问题，我们希望 了解控制的要素。 我们将使用人造酵母 染色体（YAC）绘制编码嗅觉受体的基因座。 顺式 将定义参与受体基因选择的作用DNA元素 转基因小鼠实验。 特征等位基因的实验 失活将涉及转录和DNA复制的分析 细胞系和转基因小鼠。 这些研究将阐明基因调节引导的机制 大脑中神经元的多样性的显着产生，并将 在理解大脑功能的疾病中具有价值。 而且， 哺乳动物的嗅觉神经元在生物体的整个生命中再生。 因此，了解这些神经元使用的基因表达机制是 与理解神经发育障碍特别相关。