Neural Specific Serotonin 5-HT2c Receptor

描述(由申请人提供)： 5-羟色胺受体2C亚型(5-HT2CR)是G蛋白偶联受体 这在摄食行为和紧张性抑制中起着重要作用 神经网络兴奋性。仅检测到5-HT2CR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。 了解神经递质的调节、功能和潜在的疾病关联 S-HT2CR，我们分离了人5HT2CR全长基因，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 5HT2CR 我们鉴定了另外两个cDNA变异体；一个来自于 另一个来自S-HT2CR转录本， 进行信使核糖核酸的编辑。此外，我们还发现5‘侧翼的5HL约2CR 序列可提供细胞类型特异性表达和直接转基因 对大脑的表达。几个积极和消极的监管区域 已被确认身份。特别是，我们发现了一个负面的监管区域 它似乎与多个上游元件协同调节5-HT2CR 启动子表达。在此应用程序中，我们建议进一步研究 5-HT2CR基因的调控。 这项拟议研究的中心假设是，消极的 调控元件对于正确的时间和空间表达很重要 中枢神经系统中的5-HT2CR。为了支持这一假说，两组分子 转基因方法与体外培养相结合的发育实验 提出了研究建议。首先，我们计划描述负面监管的特征 S-Hl2CR表达的重要元件及其结合蛋白。我们将首先 确认新的负元件是否调控5-HT2CR启动子 在转录水平上表达。我们将确定准确的位置 负面调控因素，并试图识别负面因素 元素结合蛋白。第二，我们将研究神经特有的和 人5-HT2CR启动子的发育性表达 负调控元件对其表达模式具有重要意义。 发展。我们计划培育出携带这种基因的转基因小鼠 5HT2CR启动子驱动的β-半乳糖苷酶基因 负调控序列。我们将分析转基因表达模式 并与内源性5-HT2CR基因进行比较。在这些基础上 分析，我希望提供一个更好的理解神经的特异性。 这种重要的神经递质受体的表达，最终导致 对中枢神经系统内部复杂的通信机制有深入的了解 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.