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.

去甲肾上腺素转运体(Net)负责限制去甲肾上腺素(NE)在大脑和交感神经系统中的活动，它通过迅速重新摄取释放的NE而发挥作用。这项建议试图确定人类网络(Hnet)基因的突变，这些突变可能导致心血管和精神疾病。立位不耐受和抑郁症患者的DNA样本将进行hnet突变的基因分型。将从血液或组织样本中提取基因组DNA，并使用针对hnet的引物，进行聚合酶链式反应和序列分析，以确定hnet编码区是否存在多态性。将通过包含hNET序列的表达载体的定点突变、[~3H]NE转运试验的转染和表现以及通过Western和放射配基结合分析进行蛋白质分析，来检查导致氨基酸替换的突变对hNET的影响。在高血压中发现的已知基因多态也将被检测其对hnet功能的影响。通过鉴定影响去甲肾上腺素功能的疾病中的hnet突变，将进一步了解去甲肾上腺素能神经传递的机制，并可以开发出针对这些疾病的选择性靶向hnet的成功治疗方法。