A calcium/calcineurin signaling cascade regulates neuronal cannabinoid receptors

钙/钙调神经磷酸酶信号级联调节神经元大麻素受体

• 批准号: 7474331
• 负责人: MARY LOU VALLANO
• 金额: $ 7.85万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7474331
• 关键词: 5' Untranslated Regions; Acute; Adolescent; Agonist; Alcohols; Animals; Antisense Oligonucleotides; Behavioral; Biochemical; Biological; Brain; CNR1 gene; Calcineurin; Calcium; Calmodulin; Cannabinoids; Cannabis; Cells; Cerebellar cortex structure; Cerebellum; Chronic; Cocaine; Consumption; Cytoplasmic Granules; Dactinomycin; Data; Development; Drug Addiction; Endocannabinoids; Future; Genes; Genetic Transcription; Heroin; Laboratories; Learning; Luciferases; Mediating; Messenger RNA; Microscopic; Molecular; Morphine; Nerve Degeneration; Neurons; Nicotine; Obesity; Oligonucleotides; Opiates; Pharmaceutical Preparations; Physiological; Plants; Polymerase Chain Reaction; Process; Promoter Regions; Proteins; Public Health; Rattus; Reporter; Repression; Rodent; Role; Seizures; Signal Transduction; Site; Structure; System; Techniques; Testing; Therapeutic; addiction; base; calcineurin phosphatase; cannabinoid receptor; cell type; excitotoxicity; inhibitor/antagonist; interest; promoter; transcription factor

DESCRIPTION (provided by applicant): This project will test the hypothesis that a Ca2+ and calmodulin-dependent phosphatase, calcineurin, regulates the transcription of type 1 cannabinoid receptors (CB1) in neurons. CB1 is the primary target for plant and endogenous cannabinoids in mammalian brain. CB1 agonists are being studied as treatments for acute and chronic neuronal degeneration, whereas antagonists appear promising to treat addictions to nicotine, alcohol, cocaine, heroin, morphine, as well as obesity. Thus there is intense interest in defining the physiological, therapeutic and pathological effects of CB1 signaling. Currently, we understand a great deal about the behavioral and pharmacological effects of cannabinoids, and CB1 structure, function and signaling. However, we know almost nothing about factors regulating the transcription CB1. My laboratory has demonstrated that a depolarization and Ca2+-dependent signaling cascade regulates expression of CB1 mRNA and protein in primary cultures of granule neurons from rodent cerebellar cortex. Our preliminary data point to primary role for calcineurin (CaN) in this process. We propose 2 specific aims. AIM 1. To test the hypothesis that Ca2+-dependent activation of CaN represses the transcription of CB1 mRNA in granule neurons from rodent cerebellar cortex. We will definitively determine the role of CaN and its downstream effector, NFAT, in CB1 transcription using a combination of biochemical, immunochemical, molecular biological and microscopic techniques. AIM 2. To test the hypothesis that CaN-dependent repression of the CB1 gene is via a promoter region containing multiple NFAT responsive sites and lying in the 5' untranslated region. PUBLIC HEALTH RELEVANCE: Cannabis is the most abused illegal drug in adolescents and chronic consumption predisposes young abusers to more serious addictions. Cannabinoid receptor (CB1) antagonists are being tested as treatments for a variety of drug addictions, including nicotine, alcohol, cocaine, and opiates. Thus, there is intense interest in defining the physiological, therapeutic and pathological effects of CB1 signaling. This project will test the hypothesis that a Ca2+ and calmodulin-dependent phosphatase, calcineurin, regulates the transcription of type 1 cannabinoid receptors (CB1) in cultured neurons from the cerebellum. Our results in cultured neurons will serve as a basis for future functional studies in animals.

描述（由申请人提供）：该项目将检验以下假设：CA2+和钙调蛋白依赖性磷酸酶钙调神经酶调节神经元中1型1型大麻素受体（CB1）的转录。 CB1是哺乳动物大脑中植物和内源性大麻素的主要靶标。正在研究CB1激动剂作为急性和慢性神经元变性的治疗方法，而拮抗剂似乎有望将成瘾治疗尼古丁，酒精，可卡因，海洛因，吗啡以及肥胖症。因此，定义CB1信号传导的生理，治疗和病理效应，人们对此非常感兴趣。目前，我们对大麻素的行为和药理作用以及CB1结构，功能和信号传导了解很多。但是，我们几乎对调节转录CB1的因素一无所知。我的实验室表明，去极化和Ca2+依赖性信号级联反应调节来自啮齿动物小脑皮质的颗粒神经元的原代培养物中CB1 mRNA和蛋白质的表达。我们的初步数据指向钙调神经酶（CAN）在此过程中的主要作用。我们提出了2个特定目标。目的1。为了测试以下假设：Ca2+依赖性激活可以抑制啮齿动物小脑皮质中颗粒神经元中CB1 mRNA的转录。我们将通过生化，免疫化学，分子生物学和微观技术的组合确定CAN及其下游效应子NFAT在CB1转录中的作用。目的2。为了测试可以依赖CB1基因的抑制作用的假设，它是通过包含多个NFAT响应位点并位于5'未翻译区域的启动子区域。公共卫生相关性：大麻是青少年中最受虐待的非法药物，长期消费使年轻的虐待者更加严重。大麻素受体（CB1）拮抗剂正在作为各种药物成瘾的处理，包括尼古丁，酒精，可卡因和阿片类药物。因此，在定义CB1信号的生理，治疗和病理效应方面，人们非常兴趣。该项目将检验以下假设：CA2+和钙调蛋白依赖性磷酸酶钙调蛋白调节来自小脑培养的神经元中1型大麻素受体（CB1）的转录。我们在培养的神经元中的结果将成为动物未来功能研究的基础。