Dopamine D1 Receptor in mouse models of primary dystonia

原发性肌张力障碍小鼠模型中的多巴胺 D1 受体

基本信息

批准号：
8583075
负责人：
MICHELLE E EHRLICH
金额：
$ 50.54万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-15 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8583075
关键词：
ATP phosphohydrolase Accounting Acute Adenosine Adenosine A2A Receptor Adenylate Cyclase Agonist Ashkenazim Basal Ganglia Base Pairing Behavior Biological Assay Brain Cells Cerebellum Clinical Collaborations Contracture Corpus striatum structure Couples Cyclic AMP DARPP DYT1 gene Data Disease Dopamine Dopamine D1 Receptor Dopamine D2 Receptor Dystonia Dystonia Musculorum Deformans Functional disorder GAG Gene GTP-Binding Proteins Genes Genetic Genotype Heterozygote Interneurons Knockout Mice Lead Letters Maps Measures Membrane Messenger RNA Molecular Molecular Profiling Motor Activity Movement Disorders Mus Muscle Mutation Nerve Degeneration Neurons Neurotransmitters Output Paper Pathway interactions Patients Phenotype Phosphorylation Population Primary Dystonias Production Property Proteins Receptor Signaling Regulation Series Signal Pathway Signal Transduction System TOR1A gene TOR1A protein Time Study Tissues TorsinA Transcriptional Regulation Tremor Tyrosine 3-Monooxygenase cholinergic cholinergic neuron chromatin immunoprecipitation comparative dopamine system early onset follower of religion Jewish in vivo mouse model mutant neurotransmission overexpression promoter protein activation public health relevance receptor reconstitution response striosome therapeutic target therapy development trafficking transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Primary torsion dystonias (PTD) are a group of movement disorders characterized by twisting muscle contractures, with dystonia as the only clinical sign (except for tremor) and in the absence of neuronal degeneration or an acquired cause. There are multiple genetic causes, with overlapping phenotypes. We have now identified a series of mutations in GNAL, encoding G?olf, in patients with early onset torsion dystonia (EOTD) who do not harbor mutations in TOR1A or THAP1. G?olf is a G protein that couples striatal dopamine D1 (D1R) and adenosine A2a (A2AR) receptors to adenylyl cyclase V. Therefore, it is expressed in striatal output medium size spiny neurons and cholinergic interneurons. Abundant evidence supports dysfunction of the basal ganglia in dystonia, although other regions, e.g. cerebellum and cortex, are also involved. Within the basal ganglia, the focus has been on the dopamine D2 receptor (D2R) and striatal cholinergic interneurons. Other than mutations in the tyrosine hydroxylase biosynthetic pathway, GNAL is the first EOTD gene that directly points to the DA signal transduction system as the origin of pathophysiology, particularly to D1R. TorsinA is a AAA-ATPase protein and Thap1 is a transcription factor. Their specific functions, however, remain enigmatic, particularly as to how their mutations result in dystonia. Therefore, the connection between G?olf and the nigrostriatal dopamine system allows for directed, comparative assays of this system in mouse "models" of the three forms of EOTD. The rationale behind these studies is that dissecting the direct effects and compensatory maladaptations in neurotransmission, particularly dopaminergic and adenosinergic, Gnal heterozygote-null mice will offer clues to pathophysiology in DYT1 (TOR1A) and DYT6 (THAP1) EOTD as well. In Specific Aim 1, it will be determined whether mutations in EOTD genes TOR1A, THAP1 and GNAL result in similar altered DA neurotransmission in the striatum as evidenced by DA level and release, G protein activity, and cAMP production. In Specific Aim 2, baseline and pharmacologically induced behavior will be analyzed in the same genotypes. The molecular counterparts of the behaviors will be assayed via measures of induction of phosphorylation of ERK and DARPP-32, following D1R, D2R, and A2AR receptor agonists and antagonists. In Specific Aim 3, RNA-seq will be performed in the Gnal+/- mouse and THAP1-C54Y knockin mouse, and compared to those in the Tor1a GAG+/-mouse (via collaboration) to identify downstream targets, particularly in neurotransmitter pathways. Identification of a final common pathway in different forms of EOTD will aid in directing discovery of therapeutic targets for this currently incurable disorder.

描述（由申请人提供）：原发性扭转肌肌张力障碍（PTD）是一组运动障碍，其特征是扭曲肌肉染色，肌张力障碍是唯一的临床症状（震颤除外），并且在没有神经元退化或获得的原因或后果原因的情况下。有多种遗传原因，具有重叠的表型。现在，我们已经确定了Gnal中的一系列突变，编码G？OLF，患有早期发作扭曲肌张力障碍（EOTD）的患者不含有TOR1A或THAP1中的突变。 g？olf是G蛋白，是伴侣纹状体多巴胺D1（D1R）和腺苷A2A（A2AR）受体与腺苷酸环化酶V。大量证据支持肌张力障碍基底神经节的功能障碍，尽管其他地区，例如小脑和皮层也参与其中。在基底神经节内，重点是多巴胺D2受体（D2R）和纹状体胆碱能中间神经元。除酪氨酸羟化酶生物合成途径中的突变外，GNAN是第一个直接指向DA信号转导系统的EOTD基因，是病理生理学的起源，特别是到D1R。 Torsina是AAA-ATPase蛋白，THAP1是转录因子。但是，它们的特定功能仍然神秘，特别是关于它们的突变如何导致肌张力障碍。因此，G？olf与鼻纹质多巴胺系统之间的连接允许在EOTD的三种形式的鼠标“模型”中对该系统进行定向，比较测定。这些研究背后的理由是，剖析神经传递的直接作用和补偿性疾病，尤其是多巴胺能和腺苷能，Gnal杂合子无效小鼠将为DYT1（TOR1A）和DYT6（THAP1）EOTD的病理生理学提供线索。在特定的目标1中，将确定EOTD基因中的突变是否会导致纹状体中的DA神经传递的类似改变，这是DA水平和释放，G蛋白活性和cAMP生产所证明的。在特定的目标2中，将在相同的基因型中分析基线和药理学诱导的行为。在D1R，D2R和A2AR受体激动剂和拮抗剂之后，将通过诱导ERK和DARPP-32的磷酸化的诱导测量来测定行为的分子对应物。在特定的AIM 3中，将在GNAL +/-小鼠和THAP1-C54Y敲蛋白小鼠中进行RNA-SEQ，并与TOR1A GAG +/-小鼠（通过协作）（通过协作）中的那些进行比较，以识别下游靶标，尤其是在神经递质途径中。以不同形式的EOTD识别最终公共途径将有助于指导目前无法治愈的疾病的治疗靶标。