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

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要新型FLPO重组酶系的靶向ni核多巴胺神经元的表征已广泛研究了黑质nigra pars commacta（SNC）的多巴胺（DA）神经元由于它们在帕金森氏病（PD）进展过程中对退化的选择性脆弱性。 DA 在PD进展过程中，紧密相邻的腹侧对接区域（VTA）的神经元在很大程度上被保留；一个现象仍然很熟悉。试图使用SNC和VTA功能的研究现代遗传工具受到重组酶线的缺乏限制核。最近的研究表明，醛中选择性地表达了醛脱氢酶1a1（Aldh1a1） SNC DA神经元和DA神经元的这种亚群经历了最广泛的变性零星PD。最近，最近已经证明了Aldh1a1能够参与A中的GABA合成途径独立于经典的GABA生物合成途径。因此，aldh1a1+ da神经元代表与PD相关的DA神经元的解剖学和神经化学上不同的人群。获取遗传获取我们在小鼠ALDH1A1基因座上开发了FLPO重组酶敲门线。使用此新颖的ALDH1A12A-FLPO系在与多巴胺转运蛋白CRE系（SLC6A3IRES-CRE）的交叉方法中将能够获得对AldH1A1+和AldH1A1-DA神经元的遗传获取。该建议旨在测试使用Aldh1a12a-FLPO选择性靶向SNC DA神经元并确定其靶标的可行性分子表型的场和多样性。我们还评估了交叉方法的实用性使用SLC6A3IRES-CRE转基因小鼠使用ALDH1A12A-FLPO指定VTA DA神经元。