GENETIC POLYMORPHISMS HUMAN NOREPINEPHRINE TRANSPORTER

人类去甲肾上腺素转运蛋白基因多态性

• 批准号: 6477041
• 负责人: MAUREEN K HAHN
• 金额: $ 4.26万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-11-16 至 2002-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6477041
• 关键词: clinical research; depression; genetic polymorphism; high throughput technology; hormone binding protein; hormone regulation /control mechanism; human subject; hypertension; membrane transport proteins; norepinephrine; nucleic acid sequence; polymerase chain reaction; postural hypotension; protein biosynthesis; protein transport; site directed mutagenesis; transfection /expression vector; western blottings

The norepinephrine transporter (NET) is responsible for limiting the actions of norepinephrine (NE) in the brain and in the sympathetic nervous system through rapid reuptake of released NE. This proposal attempts to identify mutations in the human NET (hNET) gene that may contribute to cardiovascular and psychiatric disease. DNA samples from patients with orthostatic intolerance and depression will be genotyped for hNET mutations. Genomic DNA will be isolated from blood or tissue samples, and, using primers directed against hNET, polymerase chain reaction and sequence analysis will be performed to determine if polymorphisms exist in coding region of hNET. Mutations that result in amino acid substitutions will be examined for effects on hNET through site-directed mutagenesis of an expression vector containing hNET sequences, transfection and performance of [3H]NE transport assays and protein analysis through Western and radioligand binding assays. Known polymorphisms identified in hypertension will also be examined for their effects on hNET function. Through identification of hNET mutations in diseases affecting noradrenergic function, further understanding of the mechanism of noradrenergic neurotransmission will be achieved and successful therapies for these disorders can be developed that selectively target hNET.

去甲肾上腺素转运蛋白（NET）负责限制脑中去甲肾上腺素（NE）的作用，以及通过释放NE的快速再摄取中的交感神经系统和交感神经系统中的作用。该提案试图鉴定可能导致心血管和精神病的人网基因（HNET）基因中的突变。 HNET突变的基因分型将对具有体位不耐受和抑郁症患者的DNA样品进行基因分型。基因组DNA将从血液或组织样品中分离出来，并使用针对HNET的引物，将进行聚合酶链反应和序列分析，以确定HNET编码区域中是否存在多态性。将检查导致氨基酸取代的突变，以通过位于含有HNET序列的表达向量，[3H] NE转运测定法和通过西方和放射性结合测定法的蛋白质分析的表达向量，转染和蛋白质分析的性能的诱变进行检查。在高血压中确定的已知多态性还将检查其对HNET功能的影响。通过鉴定影响去甲肾上腺素能功能的疾病中的HNET突变，将进一步了解去甲肾上腺素能神经传递的机制，并可以为这些疾病的成功疗法提供选择性靶向HNET的成功疗法。