Characterization of a novel Flpo recombinase line targeting nigral dopamine neurons

靶向黑质多巴胺神经元的新型 Flpo 重组酶系的表征

• 批准号: 9434206
• 负责人: VALINA L. DAWSON
• 金额: $ 8.18万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9434206
• 关键词: Anatomy; Autopsy; Behavioral; Cell Nucleus; Cell physiology; Cells; Communities; Data; Disease; Disease Progression; Dopamine; Electrophysiology (science); Enterobacteria phage P1 Cre recombinase; Functional disorder; Genes; Genetic; Heterogeneity; Knock-in; Knock-in Mouse; Label; Mediating; Midbrain structure; Minority; Modernization; Modification; Molecular; Mus; Neurons; Neurosciences; Parkinson Disease; Pathway interactions; Physiological; Physiology; Population; Process; Property; Reagent; Reporter; Research Personnel; Resources; Rewards; Signal Transduction; Substantia nigra structure; System; Testing; To specify; Transgenic Animals; Transgenic Mice; Ventral Tegmental Area; Viral; aldehyde dehydrogenases; design; dopamine transporter; dopaminergic neuron; experimental study; gamma-Aminobutyric Acid; intersectionality; molecular phenotype; motor control; neurochemistry; novel; pars compacta; recombinase; segregation; selective expression; single cell analysis; tool; transcriptome sequencing

Project Summary Characterization of a novel Flpo recombinase line targeting nigral dopamine neurons The dopamine (DA) neurons of the substantia nigra pars compacta (SNc) have been extensively studied because of their selective vulnerability to degeneration during Parkinson's disease (PD) progression. The DA neurons of the closely neighboring ventral tegmental area (VTA) are largely spared during PD progression; a phenomenon that remains poorly understood. Studies attempting to separate SNc and VTA function using modern genetic tools have been limited by the lack of recombinase lines that effectively separate these two nuclei. Recent studies have shown that aldehyde dehydrogenase 1a1 (Aldh1a1) is expressed selectively in SNc DA neurons and this subpopulation of DA neurons undergoes the most extensive degeneration in sporadic PD. Also it has been recently shown that Aldh1a1 is capable of participating in GABA synthesis in a pathway independent of the classic GABA biosynthetic pathway. Thus Aldh1a1+ DA neurons represent an anatomically and neurochemically distinct population of DA neurons relevant to PD. To gain genetic access to this population we have developed a Flpo recombinase knock-in line at the mouse Aldh1a1 locus. Using this novel Aldh1a12A-Flpo line in an intersectional approach with the dopamine transporter Cre line (Slc6a3IRES-Cre) we will be able to gain genetic access to both Aldh1a1+ and Aldh1a1- DA neurons. This proposal is designed to test the feasibility of using Aldh1a12A-Flpo to selectively target SNc DA neurons and to determine their target fields and diversity of molecular phenotypes. We also evaluate the utility of intersectional approaches to specify VTA DA neurons using Aldh1a12A-Flpo with Slc6a3IRES-Cre transgenic mice.

项目摘要 黑质多巴胺神经元靶向Flpo重组酶的特性研究 黑质腹侧部（substantia nigra pars rectuata，SNc）的多巴胺（dopamine，DA）神经元已被广泛研究 因为它们在帕金森病（PD）进展期间选择性地易变性。地方检察官 在PD进展过程中，紧邻腹侧被盖区（VTA）的神经元在很大程度上幸免于难; 这一现象仍然知之甚少。尝试使用以下方法分离SNc和VTA功能的研究 现代遗传工具由于缺乏有效分离这两种基因的重组酶系而受到限制 原子核。最近的研究表明，乙醛脱氢酶1a 1（Aldh 1a 1）选择性地表达于 SNc DA神经元和DA神经元的这一亚群经历了最广泛的变性， 偶发性PD。此外，最近已经表明，Aldh 1a 1能够参与GABA的合成， 途径独立于经典的GABA生物合成途径。因此，Aldh 1a 1 + DA神经元代表了一个神经元， 解剖学和神经化学上与PD相关的DA神经元的不同群体。为了获得基因 在该群体中，我们已经开发了在小鼠Aldh 1a 1基因座处的Flpo重组酶敲入系。使用此 新的Aldh 1a 12 A-Flpo系与多巴胺转运蛋白Cre系（Slc 6a 3 IRES-Cre）的交叉方法中，我们 将能够获得遗传访问Aldh 1a 1+和Aldh 1a 1- DA神经元。这项建议旨在 测试使用Aldh 1a 12 A-Flpo选择性靶向SNc DA神经元并确定其靶点的可行性 分子表型的多样性。我们还评估了交叉方法的实用性， 使用Aldh 1a 12 A-Flpo和Slc 6a 3 IRES-Cre转基因小鼠指定VTA DA神经元。