Changes in synaptic function and structure associated with chronic cocaine

与长期可卡因相关的突触功能和结构的变化

• 批准号: 7732129
• 负责人: Veronica A Alvarez
• 金额: $ 68.69万
• 依托单位: NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732129
• 关键词: AMPA Receptors; Acute; Address; Affect; Alcohol consumption; Alcohols; Animal Behavior; Animal Experimentation; Animals; Behavior; Biochemical; Biochemistry; Brain; Brain region; Calcium Signaling; Characteristics; Chromosome Pairing; Chronic; Cocaine; Cocaine Users; Daily; Dendritic Spines; Dependence; Development; Disease; Dopamine Receptor; Ethanol; Experimental Designs; Exposure to; Glutamates; Impulsivity; In Vitro; Injection of therapeutic agent; Laboratories; Lead; Long-Term Effects; Morphology; Motor Activity; Mus; N-Methyl-D-Aspartate Receptors; Neurons; Numbers; Persons; Pharmaceutical Preparations; Population; Predisposition; Prevention; Procedures; Property; Proteins; Psychiatry; Reaction Time; Receptor Signaling; Saline; Science; Score; Self Administration; Self-Administered; Signaling Molecule; Slice; Structure; Synapses; Synaptic plasticity; Techniques; Testing; Transgenic Mice; Vertebral column; Wild Type Mouse; addiction; alcohol exposure; alcohol seeking behavior; behavior test; brain tissue; cell motility; day; drug of abuse; drug seeking behavior; improved; preference; research study; size; synaptic function

To address the specific questions mentioned above, we have outline five projects and a brief experimental design summary. Project 1: How does chronic cocaine treatment affect synaptic function and morphology? Wild-type and transgenic mice will receive passive cocaine/saline administration (IP) daily for 5-30 days. This will be followed by behaivoral testing: locomotor activity and place preference tested every day with injection. We will then prepare acute brain slices / brain slice cultures and perform in vitro procedures Different techniques and in vitro procedures will be applied to determine the changes that take place in the brain of addicted animals. We will investigate: - Morphological changes: such as alterations in spine numbers, size, motility and dendritic branching. - Changes in synaptic strength, synaptic plasticity and modulation. - Biochemical changes in expression of synaptic proteins such as glutamate AMPA receptors, NMDA receptors, dopamine receptors and signaling molecules. - Changes in intracellular calcium signaling in spines and dendrites Project 2: Animals that display high impulsivity behavior show increased vulnerability to become compulsive drug seekers when chronically exposed to cocaine (Belin et al., 2008 Science 320:1352-1355). We will compare the synaptic function and the structure of neurons in different brain regions of mice that display high and low impulsivity. Wild-type and transgenic mice will undergo behavioral testing (5 choice serial reaction time task) daily for 30-40 days followed by in vitro procedures. Different techniques will be used to identify neuronal markers highly impulsive animals. We will investigate: - Morphology: spine numbers, size, motility and dendritic branching. - Biochemistry: expression levels of synaptic proteins and signaling molecules. - Calcium signaling in spines and dendrites - Synaptic strength and plasticity Project 3: Only a fraction of cocaine users become compulsive drug-takers. What is different in the brain of subjects exposed and addicted in comparison to those exposed but non-addicted? Is there a difference in synaptic function and morphology between mice that display addictive-like behaviors and mice that do not after they all have been chronically exposed to cocaine? Wild-type mice and transgenic mice will receive cocaine/saline self-administration for 30 days followed by behavioral testing to determine addictive score after 1, 10 or 30 days of last exposure. We will then perform in vitro procedures on these mice. Electrophysiological, morphological and biochemical analysis will be performed as described in project 1 and 2. Project 4: What is the effect of voluntary ethanol exposure on neuronal function and morphology in specific brain regions? What is different in the brain of animals that actively seek ethanol and those that avoid it after the chronic treatment? Wild-type and transgenic mice will undergo voluntary ethanol intake for 15-30 days followed by behavioral testing to determine degree of dependence. In vitro procedures will then be performed on brain tissue from these mice. Electrophysiological, morphological and biochemical analysis will be performed as described above.

针对上述具体问题，我们提出了五个方案和一个简短的实验设计总结。 项目1：慢性可卡因治疗如何影响突触功能和形态？ 野生型和转基因小鼠将每天接受被动可卡因/盐水施用（IP），持续5-30天。随后将进行口服测试：每天注射测试自发活动和位置偏好。 然后，我们将准备急性脑切片/脑切片培养物并进行体外程序 不同的技术和体外程序将被应用于确定成瘾动物大脑中发生的变化。我们将调查： - 形态学变化：如棘数目、大小、运动性和树突分支的改变。 - 突触强度、突触可塑性和调制的变化。 - 突触蛋白（例如谷氨酸AMPA受体、NMDA受体、多巴胺受体和信号分子）表达的生化变化。 - 棘突和树突细胞内钙信号的变化 项目二：当长期暴露于可卡因时，表现出高冲动行为的动物表现出变得强迫性药物寻求者的增加的脆弱性（贝林等人，2008 Science 320：1352-1355）。我们将比较显示高和低冲动的小鼠不同脑区的突触功能和神经元结构。 野生型和转基因小鼠将每天进行行为测试（5个选择连续反应时间任务），持续30-40天，然后进行体外程序。不同的技术将用于识别神经元标记高度冲动的动物。我们将调查： - 形态学：棘数目、大小、运动性和树突分支。 - 生物化学：突触蛋白和信号分子的表达水平。 - 棘突和树突中的钙信号 - 突触强度和可塑性 项目3：只有一小部分可卡因使用者成为强迫性吸毒者。与那些暴露但不上瘾的人相比，暴露并上瘾的受试者的大脑有什么不同？ 在慢性接触可卡因后，表现出成瘾样行为的小鼠和没有成瘾样行为的小鼠之间，突触功能和形态是否有差异？ 野生型小鼠和转基因小鼠将接受可卡因/盐水自我给药30天，然后进行行为测试，以确定最后一次暴露1、10或30天后的成瘾评分。 然后，我们将在这些小鼠身上进行体外程序。将按照项目1和2中所述进行电生理学、形态学和生化分析。 项目4：自愿乙醇暴露对特定脑区神经元功能和形态的影响是什么？主动寻求乙醇的动物和那些在长期治疗后避免乙醇的动物的大脑有什么不同？ 野生型和转基因小鼠将自愿摄入乙醇15-30天，然后进行行为测试以确定依赖程度。然后将对这些小鼠的脑组织进行体外程序。将如上所述进行电生理学、形态学和生物化学分析。