Translational analysis of functional variation in human dopamine beta?hydroxylase

人多巴胺β羟化酶功能变异的翻译分析

• 批准号: 8191158
• 负责人: Joseph F. Cubells
• 金额: $ 23.04万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8191158
• 关键词: Accounting; Address; Adrenal Medulla; Adult; Affect; Affective; Alleles; Animal Model; Antihypertensive Agents; Area; Behavior; Behavioral; Biological; Blood Pressure; Brain; Catecholamines; Cells; Chromatin; Cocaine; Data; Degenerative Disorder; Disease; Dopamine; Drug Addiction; Embryo; Embryonic Development; Environment; Enzymes; Epinephrine; Exhibits; Failure; Feedback; Gene Expression; Gene Transfer Techniques; Generations; Genes; Genetic Polymorphism; Genetic Transcription; Genetic Variation; Genomics; Goals; Growth; Health; Heterozygote; Human; Hyperactive behavior; In Vitro; Individual; Intervention; Investigation; Knock-out; Laboratories; Lead; Measurable; Mediating; Mixed Function Oxygenases; Modeling; Molecular; Mood Disorders; Mus; Neurologic; Neurotransmitters; Norepinephrine; Orthostatic Hypotension; Patients; Pattern; Peripheral; Phenotype; Physiological; Plasmids; Population Genetics; Positioning Attribute; Proteins; Psychotic Disorders; Ptosis; Pulse Pressure; Regulation; Reporter; Rest; Serum; Single Nucleotide Polymorphism; Site; Stress; Time; Transfection; Transgenes; Transgenic Mice; Transgenic Organisms; Ursidae Family; Variant; Whole Organism; Work; alcohol sensitivity; base; enzyme activity; experimental analysis; in utero; in vivo; innovation; mouse model; mutant; nervous system disorder; neurochemistry; neuromechanism; neurotransmission; noradrenergic; novel; null mutation; prenatal; public health research; relating to nervous system; research study; response

DESCRIPTION (provided by applicant): Project Summary: Dopamine 2-hydroxylase (DBH) catalyzes synthesis of the neurotransmitter norepinephrine (NE) from dopamine (DA), and thereby regulates levels of both molecules. DBH is detectable in human serum, where its activity largely reflects sequence variation at the structural locus DBH. We identified a single nucleotide polymorphism (SNP) 970 bp upstream of the transcriptional start site (-970C>T) that accounts for 30-50% of the total variance in serum DBH activity, with the "T" allele being associated with lower activity than the "C" allele, in a co-dominant pattern. Our work on a population-genetic level, and molecular experiments from another laboratory provide strong evidence that -970C>T is a true regulatory variant, affecting transcription of DBH. However, the molecular experiments suggested that the low-serum-DBH T allele associates with greater transcription. Thus, how -970C>T regulates DBH expression in vivo is not clear. The present project proposes to use BAC transgenesis, followed by crossing with Dbh -/- mice lacking endogenous DBH expression, to investigate DBH expression in brain, adrenal medulla and serum, associated with either the T or C allele in an otherwise identical human-sequence context of ~170 kb. This project will inform multiple areas of public health research, including studies of psychosis, mood disorders, drug addiction, neuro-degenerative disorders, and disorders of human blood pressure, as variation in serum DBH and/or -970C>T associates with important aspects of human phenotypes related to all of those disorders. The proposed work will produce a valuable and innovative animal model for investigation of many aspects of genetic variation in NE-mediated neurotransmission. PUBLIC HEALTH RELEVANCE: Project Narrative: Dopamine 2-hydroxylase (DBH) is an enzyme that synthesizes the neural signaling molecule norepinephrine (NE), which is in turn involved in human psychiatric, neurological and blood-pressure related disorders. A sequence variant (polymorphism) in the human DBH gene strongly regulates levels of DBH, and influences all of the foregoing human disorders. This project will put the two versions (alleles) of the human DBH gene into mice that lack their own Dbh. This will allow functional study of this important human polymorphism in a mouse model.

描述（由申请人提供）： 项目摘要：多巴胺 2-羟化酶 (DBH) 催化多巴胺 (DA) 合成神经递质去甲肾上腺素 (NE)，从而调节这两种分子的水平。 DBH 在人血清中可检测到，其活性很大程度上反映了结构位点 DBH 的序列变异。我们在转录起始位点 (-970C>T) 上游 970 bp 处发现了一个单核苷酸多态性 (SNP)，该多态性占血清 DBH 活性总变异的 30-50%，其中“T”等位基因与较低的活性相关比“C”等位基因处于共同显性模式。我们在群体遗传水平上的工作以及另一个实验室的分子实验提供了强有力的证据，证明-970C>T是一个真正的调控变异，影响DBH的转录。然而，分子实验表明低血清 DBH T 等位基因与更大的转录相关。因此，-970C>T如何调节体内DBH表达尚不清楚。本项目建议使用 BAC 转基因，然后与缺乏内源性 DBH 表达的 Dbh -/- 小鼠杂交，以研究大脑、肾上腺髓质和血清中的 DBH 表达，与其他相同的人类序列中的 T 或 C 等位基因相关~170 kb 的上下文。该项目将为公共卫生研究的多个领域提供信息，包括精神病、情绪障碍、药物成瘾、神经退行性疾病和人类血压疾病的研究，因为血清 DBH 和/或 -970C>T 的变化与重要方面相关与所有这些疾病相关的人类表型。拟议的工作将产生一个有价值且创新的动物模型，用于研究 NE 介导的神经传递遗传变异的许多方面。 公共卫生相关性： 项目叙述：多巴胺 2-羟化酶 (DBH) 是一种合成神经信号分子去甲肾上腺素 (NE) 的酶，而去甲肾上腺素又与人类精神、神经和血压相关疾病有关。人类 DBH 基因中的序列变异（多态性）强烈调节 DBH 的水平，并影响所有上述人类疾病。该项目将把人类 DBH 基因的两个版本（等位基因）植入缺乏自身 Dbh 的小鼠体内。这将允许在小鼠模型中对这一重要的人类多态性进行功能研究。