Neural Specific Serotonin 5-HT2c Receptor

描述（由申请人提供）： 5-羟色胺受体亚型2C（5-HT 2CR）是G蛋白偶联受体 在摄食行为和对大脑皮层的紧张性抑制中起着重要作用。 神经元网络兴奋性仅检测到5-HT 2CR的表达 在中枢神经系统（CNS）中，并在大脑发育过程中受到调节。 了解其调节、功能及潜在的疾病关系， S-HT 2CR，我们分离了完整的人5 HT 2CR cDNA，并对其基因进行了鉴定 结构和5'侧翼序列。除了正常的4.8-kb 5 HT 2CR外， cDNA，我们确定了另外两个cDNA变体;一个是来自一个 另一个来自S-HT 2CR转录物， 进行mRNA编辑。此外，我们发现5'侧翼5 Hl约为2CR 序列可以赋予细胞类型特异性表达和指导转基因 向大脑表达。几个积极和消极的监管区域， 被识别。特别是，我们已经定位了一个负调控区域， 它似乎与多个上游元件合作调节5-HT 2CR 启动子表达在本申请中，我们建议进一步研究 5-HT 2CR基因的调控。 这项研究的核心假设是， 调节元件对于适当的时间和空间表达是重要的， CNS中的5-HT 2CR。为了支持这一假设，两组分子 转基因方法与体外培养相结合的发育实验 研究提出。首先，我们计划将负面监管特征 元件及其对S-H12 CR表达重要的结合蛋白。我们将首先 证实新的负性元件是否调控5-HT 2CR启动子 在转录水平上表达。我们会确定 的负面调控因素，并试图确定负面的 元素结合蛋白其次，我们将研究神经特异性和 人5-HT 2CR启动子的发育表达，并检查 负调控元件对于其表达模式是重要的 发展我们计划培育出转基因小鼠， 由5 HT 2CR启动子驱动的β-半乳糖苷酶基因，有或没有 负调控序列我们将分析转基因表达模式 并与内源性5-HT 2CR基因的表达进行比较。对这些 分析，我希望能更好地了解神经特异性， 这种重要的神经递质受体表达，最终导致 深入了解CNS内复杂的通信机制 包括脉络丛，这是血脑屏障的重要组织。

DESCRIPTION (provided by applicant): The serotonin receptor subtype 2C (5-HT2CR) is a G protein-coupled receptor that plays an important role in feeding behavior and tonic inhibition of the neuronal network excitability. The expression of the 5-HT2CR is detected only in the central nervous system (CNS) and is regulated during brain development. To understand the regulation, function and potential disease association of the S-HT2CR, we isolated the complete human 5HT2CR cDNA and characterized its gene structure and 5' flanking sequence. In addition to the normal 4.8-kb 5HT2CR cDNA, we identified two other cDNA variants; one is derived from an alternatively spliced RNA and the other from the S-HT2CR transcript which undergoes mRNA editing. Also, we found that the 5' flanking 5Hl about2CR sequence can confer cell type-specific expression and direct transgene expression to the brain. Several positive and negative regulatory regions have been identified. In particular, we have located a negative regulatory region which appears to cooperate with multiple upstream elements to regulate 5-HT2CR promoter expression. In this application, we propose to further investigate the regulation of 5-HT2CR gene. The central hypothesis of this proposed research is that that the negative regulatory element is important for proper temporal and spatial expression of the 5-HT2CR in the CNS. In support of this hypothesis, two sets of molecular and developmental experiments combining transgenic approach with in vitro studies are proposed. First, we plan to characterize the negative regulatory element and its binding protein important for S-Hl2CR expression. We will first confirm whether the novel negative element regulates 5-HT2CR promoter expression at the level of transcription. We will define the precise location of the negative regulatory element and attempt to identify the negative element-binding protein. Second, we will study the neural-specific and developmental expression of the human 5-HT2CR promoter and examine if the negative regulatory element is important for its expression pattern during development. We plan to generate transgenic mice carrying the beta-galactosidase gene driven by the 5HT2CR promoter with or without the negative regulatory sequence. We will analyze the transgene expression pattern in the CNS and compare to that of the endogenous 5-HT2CR gene. Upon these analyses, I hope to provide a better understanding of the neural specificity of this important neurotransmitter receptor expression, ultimately leading to a thorough understanding of the complex communication mechanisms within the CNS including the choroid plexus, a important tissue for the blood-brain barrier.