Opiate drug abuse & HIV-induced excitotoxicity in striatal neurons

阿片类药物滥用

基本信息

批准号：
9064737
负责人：
Sylvia Fitting
金额：
$ 24.65万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-01-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9064737
关键词：
AMPA Receptors Acquired Immunodeficiency Syndrome Action Potentials Affect Behavioral Breeding CTOP Calcium Cell Culture Techniques Cells Cessation of life Coculture Techniques Cognitive deficits Complex Corpus striatum structure DRD2 gene Defect Dendrites Dendritic Spines Doctor of Philosophy Drug abuse Electrons Electrophysiology (science)Engineering Event Functional disorder Genetic Glutamates Grant HIV HIV-1 Homeostasis Image In Vitro Individual Injury Interruption Ion Channel Ions Knockout Mice Knowledge LoxP-flanked allele Mediating Membrane Membrane Potentials Mentors Mitochondria Molecular Morphine Motor Activity Mus N-Methyl-D-Aspartate Receptors N-Methylaspartate Neurocognitive Deficit Neuronal Injury Neurons Neuropathogenesis Opiates Opioid Opioid Receptor Outcome Measure Pathogenesis Pathology Pathway interactions Pharmaceutical Preparations Phase Physiological Potassium Property Proteins Research Role Site Sodium Sorting - Cell Movement Synapses Techniques Testing Therapeutic Training Varicosity Vertebral column base biophysical properties cell type cellular imaging connective tissue-activating peptide density drug abuser electrical property excitotoxicity functional loss in vivo mitochondrial membrane mouse Cre recombinase naltrindole nervous system disorder neuron loss neuronal excitability neurotoxicity norbinaltorphimine opioid abuse patch clamp prevent receptor research study skills synergism voltage

AMPA Receptors Acquired Immunodeficiency Syndrome Action Potentials Affect Behavioral Breeding CTOP Calcium Cell Culture Techniques Cells Cessation of life Coculture Techniques Cognitive deficits Complex Corpus striatum structure DRD2 gene Defect Dendrites Dendritic Spines Doctor of Philosophy Drug abuse Electrons Electrophysiology (science)Engineering Event Functional disorder Genetic Glutamates Grant HIV HIV-1 Homeostasis Image In Vitro Individual Injury Interruption Ion Channel Ions Knockout Mice Knowledge LoxP-flanked allele Mediating Membrane Membrane Potentials Mentors Mitochondria Molecular Morphine Motor Activity Mus N-Methyl-D-Aspartate Receptors N-Methylaspartate Neurocognitive Deficit Neuronal Injury Neurons Neuropathogenesis Opiates Opioid Opioid Receptor Outcome Measure Pathogenesis Pathology Pathway interactions Pharmaceutical Preparations Phase Physiological Potassium Property Proteins Research Role Site Sodium Sorting - Cell Movement Synapses Techniques Testing Therapeutic Training Varicosity Vertebral column base biophysical properties cell type cellular imaging connective tissue-activating peptide density drug abuser electrical property excitotoxicity functional loss in vivo mitochondrial membrane mouse Cre recombinase naltrindole nervous system disorder neuron loss neuronal excitability neurotoxicity norbinaltorphimine opioid abuse patch clamp prevent receptor research study skills synergism voltage

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项目摘要

Project Summary HIV-1 infected individuals who are injecting opioid drugs show increased cognitive defects and undergo an accelerated rate of progression to AIDS. Accumulating evidence suggests that opioid drug abuse intrinsically exacerbates the pathogenesis of HIV-1. We have found that neuronal death is preceded by a prolonged period of synaptic culling, functional losses, and dendritic pathology that are presumed reversible. Importantly, opioid abuse potentiates the neuropathogenesis of HIV-1 by synergistically increasing dendritic pathology (varicosity formation, beading, fragmentation, pruning), while promoting additive dendritic spine losses (plasticity). This has been verified in medium spiny neurons (MSN) of the striatum and synaptic pruning has been confirmed electron microscopically. Moreover, behavioral defects in locomotor activity are accompanied by synaptic losses and dendritic pathology in the absence of demonstrable neuron death, suggesting that sublethal neuronal injury and reduced synaptic connectivity underlie the ability of opioids to aggravate HIV-1-associated neurological disorders (HAND). While death per se is significant, the interruption of events preceding neuron death may be more strategic therapeutically. This grant will focus on the functional level of MSN by investigating the underlying physiological mechanisms of opioid ± HIV-induced excitotoxicity. It is hypothesized that Tat induces changes in the cellular homeostasis and excitability of MSN, that are exacerbated by opioid drugs through a complex sequence of events involving OR-mediated pathways. In vitro approaches are being proposed by assessing the effects of opioid drug and HIV-1 Tat-induced neurotoxicity in dissociated cortical- striatal cell cultures. Whole-cell patch-clamp recordings will be conducted in voltage- and current-clamp mode by assessing action potentials as well as sodium, potassium, AMPA, NMDA, and calcium (Ca2+) currents. The role of OR will be elucidated by applying pharmacological (OR antagonists), genetic (OR knockout mice) and silencing (silencing NMDAR) strategies to identify mechanisms underlying opioid + HIV protein interaction. To sort out whether opioids exacerbate the excitotoxic effects of Tat in the striatum via OR on MSN we will conduct experiments in vivo using two types of Cre-lox mice. Conditionally deleting OR at key sites will define the targets and associated mechanisms by which opioids exacerbate neuronal excitability (action potentials, ion channel activity, ion imaging, mitochondrial membrane potential), injury (including dendritic pathology and spine density), and behavioral defects (locomotor activity) in the striatum.

项目摘要注射阿片类药物的HIV-1感染者表明认知缺陷增加并经历加速向艾滋病的发展速率。加速证据表明阿片类药物本质上滥用药物加剧了HIV-1的发病机理。我们发现神经元死亡是长时间的重要的是，假定可逆的突触扣，功能损失和树突状病理的阿片类药物。滥用通过协同增加树突病理学增强了HIV-1的神经病作用（静脉曲张形成，串珠，碎裂，修剪），同时促进树突状脊柱损失（可塑性）。这已在纹状体的中刺神经元（MSN）中进行了验证，并已确认突触修剪电子方式。此外，突触使运动活性的行为缺陷是实现的在没有可证明的神经元死亡的情况下，损失和树突状病理神经元损伤和突触连通性降低是阿片类药物加剧HIV-1相关的能力的基础神经系统疾病（手）。虽然死亡本身很重要，但神经元之前事件的中断死亡可能是更战略理论。该赠款将重点关注MSN的功能级别研究阿片类药物±HIV诱导的兴奋性毒性的潜在生理机制。它是假设的 TAT诱导了细胞稳态的变化和MSN的兴奋性，这些变化会因卵毒素而加剧通过涉及或介导的途径的复杂事件序列的药物。体外方法正在通过评估阿片类药物和HIV-1 TAT诱导的神经毒性在解离的皮质 - 纹状体细胞培养。全细胞贴片钳记录将以电压和电流钳模式进行通过评估动作电位以及钠，钾，AMPA，NMDA和钙（Ca2+）电流。这通过应用药物（或拮抗剂），遗传（或敲除小鼠）和沉默（沉默NMDAR）策略，以识别阿片类药物 + HIV蛋白相互作用的机制。到弄清阿片类药物是否加剧了TAT通过MSN或MSN的纹状体中TAT的兴奋性毒性作用使用两种类型的Cre-lox小鼠在体内进行实验。有条件删除或在关键站点定义阿片类药物加剧神经元兴奋性的目标和相关机制（动作电位，离子通道活性，离子成像，线粒体膜电位），损伤（包括树突病理学和脊柱密度）和纹状体中的行为缺陷（运动活性）。