Multiphoton Monoamine Imaging of Serotonin Neuron Function

血清素神经元功能的多光子单胺成像

• 批准号: 7570447
• 负责人: EDWIN S LEVITAN
• 金额: $ 14.48万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7570447
• 关键词: Action Potentials; Address; Affect; Amphetamines; Anxiety; Axon; Biological Assay; Brain; Cell Nucleus; Cocaine; Cultured Cells; Dendrites; Development; Drug abuse; Ensure; Grant; Guidelines; Image; Lasers; Light; Mental Depression; Methods; Moods; Neurons; Neurotransmitters; Optical Methods; Optics; Pharmaceutical Preparations; Physiologic pulse; Physiological; Property; Public Health; Pulse taking; Regulation; Rewards; Sapphire; Serotonin; Site; Slice; Stimulus; System; Techniques; Tissues; Varicosity; Vesicle; addiction; analog; dorsal raphe nucleus; drug of abuse; ecstasy; monoamine; neuronal cell body; patch clamp; raphe nuclei; research study; uptake

DESCRIPTION (provided by applicant): Serotonin neurons of the raphe nuclei control mood and are sensitive to drugs of abuse such as the amphetamine MDMA (3,4-Methylenedioxymethamphetamine, Ecstasy) and cocaine. Because serotonin release by axon-derived varicosities, cell bodies and dendrites cannot be assayed and compared directly in intact tissue, the contribution of each cellular compartment to release evoked by electrical activity and drugs has not been quantified. Likewise, the properties of release sites near a dorsal raphe nucleus serotonin neuron that arise from that neuron, from neighboring neurons in the same nucleus and projections from other raphe nuclei have not been determined with current methods. Therefore, we have been developing a new optical technique for studying serotonin neurons in brain slices. Multiphoton monoamine imaging (MMI) utilizes a fluorescent serotonin analog that is excited by 725 nm pulsed infrared light from Ti:sapphire laser. We have demonstrated that MMI detects uptake and depolarization-evoked release from serotonin neuron cell bodies and varicosities in dorsal raphe nucleus brain slices. Here, experiments are presented to establish the utility of MMI for studying release in brain slices induced by relevant stimuli (action potentials and MDMA) at identified compartments (axonal boutons, dendrites and cell bodies) of serotonin neurons. This further development will ensure that MMI will be able to address fundamental questions concerning the effects of abused drugs on serotonin neurons. PUBLIC HEALTH RELEVANCE: Drug abuse continues to be a major public health problem. It is known that the neurotransmitter serotonin affects mood (e.g. depression, anxiety) and dopaminergic reward systems involved in addiction. This CEBRA application will further develop a new optical method called Multiphoton Monoamine Imaging for studying transmitter release by serotonin neurons.

描述（由申请人提供）：Raphe核控制情绪的5-羟色胺神经元，对滥用药物（例如苯丙胺MDMA（3,4-甲基二甲基甲基甲基甲基苯丙胺，狂喜）和可卡因）敏感。由于轴突衍生的静脉曲张释放5-羟色胺，因此无法分析细胞体和树突，并直接在完整的组织中进行比较，因此尚未量化每个细胞室对通过电活动和药物启用的释放释放的贡献。同样，尚未使用当前方法确定来自该神经元的背侧raphe核神经元的释放位点的性质，以及来自相同核中相邻神经元的相邻神经元以及来自其他raphe核的投影的性质。因此，我们一直在开发一种用于研究脑切片中5-羟色胺神经元的新光学技术。多光子单胺成像（MMI）使用了荧光5-羟色胺类似物，该类似物被Ti：Sapphire Laser的725 nm脉冲红外光激发。我们已经证明，MMI检测到从5-羟色胺神经元细胞体中的摄取和去极化诱发的释放以及背侧raphe核脑切片中的静脉曲张。在这里，提出了实验，以建立MMI在识别出的脑瘤神经元的隔室（轴突装饰物，树突和细胞体）对相关刺激（动作电位和MDMA）诱导的脑切片中的释放效用。这一进一步的发展将确保MMI能够解决有关滥用药物对5-羟色胺神经元影响的基本问题。公共卫生相关性：滥用药物仍然是一个重大的公共卫生问题。众所周知，神经递质5-羟色胺会影响成瘾中涉及的情绪（例如抑郁症，焦虑）和多巴胺能奖励系统。该CEBRA应用将进一步开发一种新的光学方法，称为多光子单胺成像，用于研究5-羟色胺神经元释放发射机。