Generation of cell-specific tools to determine the role of spinophilin in regulating pathological responses to psychostimulant drugs of abuse

生成细胞特异性工具来确定亲旋蛋白在调节精神兴奋剂滥用的病理反应中的作用

• 批准号: 9132453
• 负责人: Anthony J. Baucum
• 金额: $ 18.47万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9132453
• 关键词: Amphetamines; Animals; Applications Grants; Area; Axon; Behavior; Behavioral; Benchmarking; Biochemical; Biological; Biological Assay; Brain; CRISPR/Cas technology; Cells; Chronic; Cocaine; Corpus striatum structure; Coupled; Cyclic AMP-Dependent Protein Kinases; DRD2 gene; Dendritic Spines; Disease; Dopamine; Dopamine D1 Receptor; Dopamine D2 Receptor; Dose; Drug toxicity; Electrons; Ensure; Exons; Exposure to; G-Protein-Coupled Receptors; Generations; Genes; Glutamates; Goals; Human; Illicit Drugs; Ion Channel; Knock-out; Knockout Mice; Lead; LoxP-flanked allele; Mediating; Methamphetamine; Modeling; Molecular; Movement; Mus; Neurons; Neurosciences; Nucleus Accumbens; Parkinson Disease; Pathology; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Population; Protein phosphatase; Proteins; Regimen; Regulation; Research; Risk; Rodent; Role; Signal Transduction; Site; Substantia nigra structure; Synapses; Tamoxifen; Testing; Toxic effect; Transgenic Mice; Ventral Striatum; Vertebral column; addiction; behavioral sensitization; cell type; cocaine exposure; density; dopaminergic neuron; drug of abuse; improved; interest; knockout animal; methamphetamine abuse; novel; postsynaptic; programs; protein expression; protein function; psychostimulant; public health relevance; recombinase; response; scaffold; spinophilin; success; synaptic function; targeted treatment; tool

 DESCRIPTION: The brain is made up of many different types of neurons. For example, one class of neuron in the striatum is called the medium spiny neuron (MSN). These spiny neurons are so called because they contain dendritic spines that form connections, called synapses, with glutamate-containing axons from the cortex and dopamine containing axons from the substantia nigra. The dendritic spines on the MSNs contain signaling and scaffolding molecules in an electron dense area called the postsynaptic density. Psychostimulant drugs of abuse act to increase dopamine release from nigral dopaminergic neurons, which has downstream effects on proteins that are localized to MSN dendritic spines. One protein, spinophilin, is increased upon chronic exposure to psychostimulant drugs of abuse. Striatal MSNs can be subdivided into two classes, direct and indirect pathway neurons. Currently there are very few ways to evaluate the function of synaptic proteins in the different classes of MSNs. This is important because the two classes of MSNs have different responses to psychostimulant exposure. The goal of our proposal is to determine the function of spinophilin in the two classes of MSNs in regulating responses to chronic non-toxic and toxic regimens of psychomotor stimulants such as cocaine and amphetamines. Determining the cell-specific role of spinophilin in mediating pathologies associated with these drugs of abuse will identify new pathways that can be targeted for treatment of addiction and disorders associated with long-term methamphetamine abuse, such as Parkinson disease (PD). In this proposal, we hypothesize that spinophilin plays a unique role in the two MSN populations in mediating psychostimulant-induced pathologies. This hypothesis will be tested using the following specific aims: Aim 1. (R21). Generate and validate mouse lines expressing tamoxifen-inducible, iCre in direct and indirect pathway MSNs. We will generate mouse lines that express a tamoxifen-inducible, improved Cre (iCre) recombinase in direct or indirect pathway MSNs. Aim 2. (R21). Generate and validate floxed spinophilin (Spinofl/fl) mice. We will generate a novel transgenic mouse line that will be crossed with the iCre animals generated in aim 1 to create MSN-specific spinophilin KO animals. Aim 3. (R33). Determine the role of spinophilin in direct and indirect pathway MSNs on psychostimulant sensitization. We will use mice created in aims 1 and 2 to determine the function of spinophilin in the two MSN populations on the regulation of pathologies associated with psychostimulant-induced sensitization Aim 4. (R33). Determine the role of spinophilin in direct and indirect pathway MSNs on METH toxicity. We will use mice created in aims 1 and 2 to determine the function of spinophilin in the two MSN populations on the regulation of pathologies associated with methamphetamine-induced toxicity.

 描述：大脑由许多不同类型的神经元组成。例如，纹状体中的一类神经元称为中刺神经元（MSN）。之所以称呼这些棘神经元，是因为它们包含形成连接的树突状刺，称为突触，并带有来自皮质的含谷氨酸的轴突，而多巴胺中含有来自黑质的轴突的多巴胺。 MSN上的树突状刺包含信号传导和脚手架分子，称为突触后密度。精神刺激药物的滥用药物可以增加果膜多巴胺能神经元释放多巴胺，该神经元对局部属于MSN树突状棘的蛋白质具有下游作用。长期暴露于精神刺激药物的滥用药物后，一种蛋白质，旋转蛋白会增加。纹状体MSN可以细分为两类，直接和间接途径神经元。当前，在不同类别的MSN类别中评估突触蛋白的功能的方法很少。这很重要，因为这两类MSN对心理刺激暴露有不同的反应。我们建议的目的是确定三类MSN中自旋素的功能，以确定对可卡因和苯丙胺等精神运动刺激剂的慢性无毒和毒性方案的反应。确定自旋蛋白在与这些滥用药物相关的介导病理中的细胞特异性作用将确定可用于治疗成瘾和与长期甲基苯丙胺滥用相关的疾病的新途径，例如帕金森氏病（PD）。在此提案中，我们假设自旋磷酸在两个MSN群体中起着独特的作用，在介导精神刺激剂诱导的病理学中起着独特的作用。该假设将使用以下特定目的进行检验：AIM 1。（R21）。在直接和间接途径MSN中生成和验证表达他莫昔芬诱导的ICRE的小鼠系。我们将生成在直接或间接途径MSN中表达他莫昔芬可诱导的，改进的CRE（ICRE）重组酶的小鼠系。目标2。（R21）。产生并验证floxed Spinophilin（Spinefl/fl）小鼠。我们将生成一条新型的转基因小鼠系，该系列将与AIM 1中产生的ICRE动物交叉，以创建MSN特异性自旋素KO动物。目标3。（R33）。确定自旋素在直接和间接途径MSN中的作用在心理刺激敏感性中。我们将使用在目标1和2中创建的小鼠来确定两个MSN群体中自旋素在与精神刺激诱导的敏感性目标相关的病理学调节中的功能。4。（R33）。确定自旋素在直接和间接途径MSN中的作用在甲基毒性中。我们将使用在目标1和2中产生的小鼠来确定两个MSN群体中自旋素对与甲基苯丙胺诱导的毒性相关的病理学调节的功能。