Spinophilin Signaling in the Striatum

纹状体中的 Spinphilin 信号传导

• 批准号: 8811481
• 负责人: Anthony J. Baucum
• 金额: $ 13.87万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8811481
• 关键词: 1 year old; Accounting; Address; Affect; Aging; Animal Disease Models; Animal Model; Animals; Antibodies; Autopsy; Bacterial Artificial Chromosomes; Binding Proteins; Biochemical; Biochemistry; Calcium; Cells; Cellular Morphology; Complex; Corpus striatum structure; Coupled; Data; Dendritic Spines; Development; Disease; Dopamine; Dopamine D1 Receptor; Dopamine D2 Receptor; Drug Targeting; Dynorphins; F-Actin; Globus Pallidus; Glutamates; Imagery; Immunoprecipitation; K-Series Research Career Programs; Knock-out; Lead; Learning; Mass Spectrum Analysis; Medial; Mediating; Mental disorders; Molecular; Monkeys; Morphology; Motor; Movement; Mus; Neurodegenerative Disorders; Neurons; Neurotransmitters; Parkinson Disease; Parkinsonian Disorders; Pathway interactions; Patients; Phosphorylation; Play; Protein Binding; Protein Dephosphorylation; Protein Isoforms; Protein phosphatase; Proteins; Proteomics; Research Design; Rodent; Role; Scaffolding Protein; Signal Transduction; Specificity; Structure; Substance P; Substantia nigra structure; Synapses; Techniques; Technology; Testing; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Vertebral column; age related; base; calmodulin-dependent protein kinase II; density; differential expression; follow-up; in vivo; innovation; inorganic phosphate; insight; intercellular communication; nervous system disorder; neurotransmission; normal aging; novel; postnatal; protein protein interaction; research study; response; spinophilin; synaptic function; tool; tool development

DESCRIPTION (provided by applicant): Parkinson disease (PD) is proximally caused by degeneration of dopamine containing neurons that project from the Substantial Nigra to the striatum. These neurons form synaptic connections with structures termed dendritic spines that reside on two subpopulations of the medium spiny neuron (MSN) that express either D1 dopamine receptors or D2 dopamine receptors (D1R and D2R, respectively). The dendritic spines also receive glutamatergic synaptic inputs from the cortex that activates calcium-dependent signaling in spines. Thus, the release of dopamine differentially modulates the actions of glutamate within the dendritic spines of D1R- and D2R-containing MSNs. Dopamine-depletion in PD patients and in parkinsonian animal models results in loss of dendritic spines from striatal MSNs; however, spine loss is restricted to D2R-containing MSNs in short-term rodent studies. Moreover, emerging data from several labs indicate that corticostriatal synapses onto D1R- and D2R-containing MSNs are differentially regulated by dopamine and other neurotransmitters. However, very little is known about the biochemical differences between striatal MSN subpopulations that presumably account for these differences. This project investigates the roles of spinophilin in D1R- and D2R-containing MSNs. Spinophilin is a scaffolding protein that binds protein phosphates 1 (PP1), F-actin, and several other proteins involved in regulating cell signaling and morphology. The global knockout of spinophilin disrupts corticostriatal synaptic function in both D1R- and D2R-containing MSNs, and also affects the morphology of striatal MSNs in an age- dependent manner. My recent studies showed that dopamine depletion enhances the interaction of spinophilin with PP1¿1; presumably modulating the dephosphorylation of other associated dendritic proteins. In order to identify potential substrates of the spinophilin-PP1 complex, I performed a proteomics screen, identifying multiple spinophilin-associated proteins (SpAPs) in normal, mature striatum that are known to regulate cell morphology, including CaMKII. Previous studies in this lab showed that dopamine depletion leads to hyper- phosphorylation of CaMKII. In this career development award I will develop innovative transgenic molecular tools to address my over-arching hypothesis that: dopamine depletion and aging differentially regulate spinophilin-dependent signaling in striatal MSN subtypes. Two aims will begin to test this hypothesis: Aim 1 will test the hypothesis that dopamine depletion alters the spinophilin interaction network in an age-dependent manner. Aim 2 will use novel transgenic animals expressing differentially tagged forms of spinophilin in D1R- or D2R- containing MSNs to test the hypothesis that dopamine depletion alters the spinophilin interaction network in a cell-specific manner. These studies will greatly enhance our understanding of spinophilin-mediated striatal signaling in animal models of PD and will inform potential drug targets for this disorder.

描述（由应用提供）：帕金森氏病（PD）是由含有多巴胺的变性引起的，这些神经元含有从实质性nigra到纹状体的神经元的变性。这些神经元与称为树突状棘的结构形成突触连接，这些结构位于中等棘神经元（MSN）的两个亚群，这些神经元（MSN）分别表达D1多巴胺受体或D2多巴胺受体（D1R和D2R）。树突状刺还从皮质中接收谷氨酸能突触输入，从而激活棘突中的钙依赖性信号。这是多巴胺的释放对D1R-和D2R的MSN的树突状棘的作用进行了不同的调节。 PD患者和帕金森氏动物模型中的多巴胺 - 止动物导致纹状体MSN的树突状棘损失；但是，在短期啮齿动物研究中，脊柱损失仅限于含D2R的MSN。此外，来自多个实验室的新兴数据表明，在含D1R和D2R的MSN上的皮质纹状体突触受多巴胺和其他神经递质的差异调节。但是，对于可能解释这些差异的纹状体MSN亚群之间的生化差异知之甚少。该项目调查了酚磷脂在含D1R和D2R的MSN中的作用。 Spinophilin是一种结合蛋白质磷酸盐1（PP1），F-肌动蛋白和其他参与调节细胞信号传导和形态的其他蛋白质的支架蛋白。自旋素的全球敲除在含D1R和D2R的MSN中破坏皮质纹状体突触功能，并以年龄依赖性方式影响纹状体MSN的形态。我最近的研究表明，多巴胺的耗竭增强了自旋磷脂与pp1¿1的相互作用。大概调节了其他相关的树突状蛋白的去磷酸化。为了鉴定自旋蛋白-PP1复合物的潜在底物，我进行了蛋白质组学筛查，在正常的成熟纹状体中鉴定了多种自旋素相关蛋白（空位），这些蛋白质（spaps）已知可以调节包括CAMKII在内的细胞形态。该实验室的先前研究表明，多巴胺的耗竭会导致CAMKII的过度磷酸化。在这个职业发展奖中，我将开发创新的转基因分子工具，以解决我的架构过度假设：多巴胺耗竭和衰老对纹状体MSN亚型中的旋转蛋白依赖性信号传导的调节不同。两个目标将开始检验这一假设：AIM 1将检验以下假设：多巴胺耗竭以年龄依赖性方式改变自旋素相互作用网络。 AIM 2将使用新型的转基因动物，这些动物在含有MSN的D1R-或D2R-中表达不同标记的自旋素形式，以测试多巴胺耗竭以细胞特异性方式改变Spinophilin相互作用网络的假说。这些研究将极大地增强我们对PD动物模型中自旋蛋白介导的纹状体信号传导的理解，并将为该疾病的潜在药物靶标提供信息。