Spinophilin Signaling in the Striatum

纹状体中的 Spinphilin 信号传导

基本信息

批准号：
9041042
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
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9041042
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 Electrophysiology (science)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 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

项目摘要

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（由申请人提供）：帕金森病（PD）近端是由从黑质投射到纹状体的含有多巴胺的神经元变性引起的，这些神经元与位于中棘的两个亚群上的称为树突棘的结构形成突触连接。表达 D1 多巴胺受体或 D2 多巴胺受体（分别为 D1R 和 D2R）的神经元 (MSN)。树突棘还接收来自皮质的谷氨酸突触输入，激活树突棘中的钙依赖性信号传导，因此，多巴胺的释放差异性地调节含有 D1R 和 D2R 的 MSN 的树突棘中谷氨酸的作用。在帕金森病动物模型中，纹状体 MSN 的树突棘缺失；然而，树突棘缺失是此外，来自多个实验室的新数据表明，含有 D1R 和 D2R 的 MSN 上的皮质纹状体突触受到多巴胺和其他神经递质的差异调节。纹状体 MSN 亚群之间的差异可能是造成这些差异的原因。该项目研究了 Spinophilin 在 D1R- 和 D1R- 中的作用。含有 D2R 的 MSN 是一种支架蛋白，可结合磷酸盐 1 (PP1)、F-肌动蛋白和其他几种参与调节细胞信号传导和形态的蛋白质。我最近的研究表明，多巴胺消耗会以年龄依赖性方式影响纹状体 MSN 的形态。增强亲旋蛋白与 PP1¿1 的相互作用；可能调节其他相关树突蛋白的去磷酸化为了鉴定亲旋蛋白-PP1 复合物的潜在底物，我进行了蛋白质组学筛选，鉴定了正常情况下的多个亲旋蛋白相关蛋白 (SpAP)。已知可调节细胞形态的成熟纹状体，包括 CaMKII，该实验室之前的研究表明，多巴胺耗竭会导致细胞过度磷酸化。 CaMKII。在这个职业发展奖中，我将开发创新的转基因分子工具来解决我的总体假设：多巴胺消耗和衰老对纹状体 MSN 亚型中的亲旋蛋白依赖性信号传导有不同的调节：目标 1 将开始检验这一假设。测试多巴胺耗竭以年龄依赖性方式改变亲旋蛋白相互作用网络的假设，目标 2 将使用表达不同标记形式的亲旋蛋白的新型转基因动物。含有 D1R 或 D2R 的 MSN 来测试多巴胺耗竭以细胞特异性方式改变亲旋蛋白相互作用网络的假设，这些研究将极大地增强我们对 PD 动物模型中亲旋蛋白介导的纹状体信号传导的理解，并将为潜在的药物靶点提供信息。对于这种疾病。