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: 7674555
• 负责人: EDWIN S LEVITAN
• 金额: $ 14.58万
• 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 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7674555
• 关键词: Action Potentials; Address; Affect; Amphetamines; Anxiety; Axon; Biological Assay; Brain; Cell Culture Techniques; Cell Nucleus; Cocaine; Dendrites; Development; Drug abuse; Ensure; Grant; Guidelines; Image; Lasers; Light; Methods; Moods; Neurons; Neurotransmitters; Optical Methods; Optics; Pharmaceutical Preparations; Physiologic pulse; Physiological; Property; Public Health; Regulation; Rewards; Sapphire; Serotonin; Site; Slice; Stimulus; System; Techniques; Tissues; Varicosity; Vesicle; addiction; analog; depression; dorsal raphe nucleus; drug of abuse; ecstasy; monoamine; neuronal cell body; patch clamp; public health relevance; 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(申请人提供)：中缝核团的5-羟色胺神经元控制情绪，对滥用药物如苯丙胺MDMA(3，4-亚甲基二氧甲基苯丙胺，摇头丸)和可卡因敏感。由于轴突来源的静脉曲张、细胞体和树突释放的5-羟色胺不能在完整的组织中直接测定和比较，因此每个细胞隔室对电活动和药物引起的释放的贡献还没有被量化。同样，中缝背核5-羟色胺神经元附近的释放部位、同一核中相邻神经元和其他中缝核团的投射的性质还没有用目前的方法确定。因此，我们一直在开发一种新的光学技术来研究脑片中的5-羟色胺神经元。多光子单胺成像(MMI)利用了一种由钛宝石激光发出的725 nm脉冲红外光激发的荧光5-羟色胺类似物。我们已经证明，MMI可以检测到5-羟色胺神经元胞体的摄取和去极化诱发的释放，以及中缝背核脑片的突触。在此，实验旨在建立MMI用于研究相关刺激(动作电位和MDMA)在5-羟色胺神经元已识别的隔室(轴突、树突和细胞体)引起的脑片释放的有效性。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。与公共卫生的相关性：药物滥用仍然是一个主要的公共卫生问题。众所周知，神经递质5-羟色胺影响情绪(如抑郁、焦虑)和与成瘾有关的多巴胺能奖赏系统。CEBRA的这一应用将进一步开发一种名为多光子单胺成像的新光学方法，用于研究5-羟色胺神经元释放的递质。