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（由申请人提供）：中缝核的血清素神经元控制情绪，并对滥用药物敏感，例如苯丙胺 MDMA（3,4-亚甲二氧基甲基苯丙胺，摇头丸）和可卡因。由于轴突衍生的静脉曲张、细胞体和树突释放的血清素无法在完整组织中直接进行测定和比较，因此每个细胞区室对电活动和药物引起的释放的贡献尚未量化。同样，中缝背核血清素神经元附近的释放位点的特性尚未用目前的方法确定，这些释放位点源自该神经元、同一核中的邻近神经元以及来自其他中缝核的投射。因此，我们一直在开发一种新的光学技术来研究脑切片中的血清素神经元。多光子单胺成像 (MMI) 利用荧光血清素类似物，该类似物由钛蓝宝石激光器发出的 725 nm 脉冲红外光激发。我们已经证明，MMI 可以检测血清素神经元细胞体的摄取和去极化诱发的释放以及中缝背核脑切片中的静脉曲张。在这里，实验旨在确定 MMI 在研究血清素神经元的相关刺激（动作电位和 MDMA）所诱导的脑切片中的释放方面的效用。这一进一步的发展将确保 MMI 能够解决有关滥用药物对血清素神经元影响的基本问题。公共卫生相关性：药物滥用仍然是一个主要的公共卫生问题。众所周知，神经递质血清素会影响情绪（例如抑郁、焦虑）和与成瘾有关的多巴胺能奖励系统。该 CEBRA 应用将进一步开发一种称为多光子单胺成像的新光学方法，用于研究血清素神经元的递质释放。