Neurochemical Basis of Opiate Addiction

阿片成瘾的神经化学基础

基本信息

批准号：
8383459
负责人：
PING-YEE LAW
金额：
$ 15.94万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
1979
资助国家：
美国
起止时间：
1979-01-01 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8383459
关键词：
Absence of pain sensation Accounting Address Adenylate Cyclase Agonist Area Arrestins Behavioral Biochemical Brain region Cells Chronic Complex Dependence Development Dominant-Negative Mutation Drug Exposure Drug effect disorder Embryo Etorphine Family Fibroblasts Fluorescence Future G-Protein-Coupled Receptors GTP-Binding Proteins Goals Gray unit of radiation dose In Vitro Knockout Mice Laboratories Link Mediating Mediator of activation protein Modeling Molecular Morphine Mus N-Methyl-D-Aspartate Receptors Naloxone Opiate Addiction Opiates Opioid Opioid Receptor Pathway interactions Peptides Pharmaceutical Preparations Phosphorylation Phosphotransferases Process Progress Reports Protein Kinase Protein Kinase C Proteins Proto-Oncogene Proteins c-raf Receptor Signaling Recruitment Activity Research Rhodopsin Role Signal Pathway Signal Transduction Small Interfering RNA Spinal Subfamily lentivirinae Viral Virus Withdrawal base desensitization design drug development in vivo inhibitor/antagonist member mutant neurochemistry opiate tolerance receptor response scaffold src-Family Kinases

项目摘要

DESCRIPTION (provided by applicant): For years, many laboratories, including ours, have focused on elucidating the molecular and cellular basis for opiate tolerance and withdrawal. Regardless of the hypotheses, one constant is that chronic drug effects are initiated by receptor signaling. Since the ?-opioid receptor (OPRM1) is a member of the rhodopsin family of G protein-coupled receptors (GPCR), the model for GPCR desensitization involving the G protein-receptor kinase (GRK) and ?-arrestin was applied to the ?-opioid receptor (OPRM1) to account for the chronic drug effect. However, other signaling pathways, such as N-methyl-D-aspartate (NMDA) receptor, protein kinase C (PKC) and even ?-opioid receptor (OPRD1), have been implicated in morphine tolerance development. The involvement of multiple protein kinases in chronic morphine effect is best exemplified by our recent observations that both Src kinase and Raf-1 kinase participate in adenyly cyclase (AC) superactivation after chronic drug treatment. These observations and others have led us to propose the hypothesis that recruitment of protein kinases by the agonist-OPRM1 complex will determine the pathway selected for cellular adaptational processes, such as opiate tolerance and withdrawal. Thus, the goals of our proposed studies are to determine whether there is an agonist-selective mechanism (i.e., protein kinase-dependent) in opiate tolerance development and whether PKC is involved in the blunting of in vivo morphine and not other agonist actions. In addition, the roles of the Src/Raf-1 kinase signaling cascade and the phosphorylation of OPRM1 in AC superactivation will be elucidated. The significance of our in vitro observations will be validated with the proposed in vivo studies. By using the approach of viral delivery of the wild type or mutant OPRM1, wild type or phosphorylation minus mutant of the PTX-insensitive Gi/o ?-subunits, or siRNA constructs to regulate the protein kinases involved, into vlPAG area of OPRM1-/- mice or double knockout mice of OPRM1 and ?Arrestin2, we will address our hypothesis with 2 specific aims: (1) To demonstrate that PKC mediates the in vivo tolerance development to morphine; (2) To delineate the pathway involved in Src-mediated OPRM1-directed AC superactivation and linking AC superactivation to naloxone precipitated withdrawal signs. We anticipate that we will demonstrate that the agonist-selective pathway in opiate tolerance development, i.e., both ?Arr and PKC pathways, are involved in tolerance development and that, by controlling the Src kinase activity within the OPRM1 signaling complex, the phosphorylation of AC by Raf-1 activated by Src leads to some if not all of the withdrawal signs observed during naloxone-precipitated withdrawal in mice chronically treated with an opioid agonist. Our proposed studies will link the action of the various protein kinases recruited and activated by OPRM1 in the behavioral responses to the chronic drug treatment.

描述（由申请人提供）：多年来，许多实验室（包括我们的实验室）一直集中在阐明分子和细胞基础上的鸦片耐受性和戒断。无论假设如何，一个常数是通过受体信号引发慢性药物作用。既然是e-opioid受体（OPRM1），是G蛋白偶联受体的Rhopopsin家族（GPCR）的成员，GPCR脱敏的模型涉及G蛋白受体激酶（GRK）和？arrestin，以构成？ - 核酸含量为？ - 核酸含量（OPRM1），以构成chrmopioid受体（OPRM1）。但是，其他信号通路，例如N-甲基-D-天冬氨酸（NMDA）受体，蛋白激酶C（PKC），甚至是？opioid受体（OPRD1），已与吗啡耐受性发展有关。我们最近观察到的，即SRC激酶和RAF-1激酶参与慢性药物治疗后参与腺苷（AC）超活化，最好用多种蛋白激酶参与慢性吗啡作用。这些观察结果和其他观察结果使我们提出了这样的假设：Agonist-Oprm1复合物对蛋白激酶的募集将确定选择用于细胞适应过程的途径，例如阿片类药物的耐受性和戒断。因此，我们提出的研究的目标是确定在阿片类药物耐受性发展中是否存在激动剂选择性机制（即蛋白激酶依赖性），以及PKC是否参与体内吗啡的钝化和其他激动剂作用。此外，将阐明SRC/RAF-1激酶信号级联和OPRM1磷酸化的作用。我们的体外观察的意义将通过拟议的体内研究来验证。通过使用野生型或突变体OPRM1的病毒输送方法，对PTX不敏感的Gi/O？ - subunits或siRNA构建的野生型或磷酸化减去突变体，以调节所涉及的蛋白激酶，以调节OPRM1 - / - 小鼠或双敲门型小鼠的VLPAG面积（以下是oprm1 and oprm1 and oprm1 and oprm1 and？证明PKC介导了对吗啡的体内耐受性发展。（2）描述参与SRC介导的OPRM1指导的AC超级激活的途径，并将AC超激活与纳洛酮连接到沉淀的戒断迹象。 We anticipate that we will demonstrate that the agonist-selective pathway in opiate tolerance development, i.e., both ?Arr and PKC pathways, are involved in tolerance development and that, by controlling the Src kinase activity within the OPRM1 signaling complex, the phosphorylation of AC by Raf-1 activated by Src leads to some if not all of the withdrawal signs observed during naloxone-precipitated withdrawal in长期用阿片类药物激动剂治疗的小鼠。我们提出的研究将将OPRM1募集和激活的各种蛋白激酶在对慢性药物治疗的行为反应中的作用联系起来。