The role L-type calcium channels (LTCCs) in regulating dopaminergic activity during cocaine abstinence

L型钙通道（LTCC）在可卡因戒断期间调节多巴胺能活性中的作用

• 批准号: 10198057
• 负责人: Nour Al-muhtasib
• 金额: $ 4万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10198057
• 关键词: Abstinence; Address; Automobile Driving; Birth; Brain; CNR1 gene; CRISPR/Cas technology; Calcium; Calcium-Binding Proteins; Cells; Cocaine; Cre driver; Data; Dendrites; Dendritic Spines; Development; Disease model; Electrophysiology (science); Endocannabinoids; Epilepsy; Equilibrium; Functional disorder; Glutamates; Goals; Hippocampus (Brain); In Situ Hybridization; Inhibitory Synapse; Interneurons; Investigational Therapies; Knowledge; L-Type Calcium Channels; Laser Scanning Microscopy; Lead; Long-Term Depression; LoxP-flanked allele; Mediating; Mental disorders; Molecular; Mus; Neocortex; Neurodevelopmental Disorder; Neurons; Neuropeptides; Output; Parvalbumins; Pathway interactions; Peptides; Pharmacology; Pharmacotherapy; Physiological; Play; Population; Regulation; Research; Role; Schizophrenia; Serotonin; Signal Transduction; Somatostatin; Structure; Synapses; Synaptic plasticity; Transgenic Mice; Vasoactive Intestinal Peptide; Viral; area striata; autism spectrum disorder; cell type; cellular targeting; endocannabinoid signaling; experimental study; gamma-Aminobutyric Acid; hippocampal pyramidal neuron; insight; mature animal; migration; nerve supply; nervous system disorder; neuropsychiatric disorder; novel; operation; optogenetics; postsynaptic; presynaptic; receptor; receptor expression; synaptic depression; synaptogenesis; targeted treatment; tool; transmission process; two-photon

The overall goal of this proposal is to investigate the role of endocannabinoid (ECB) signaling in regulating distinct cortical inhibitory synapses. Physiological brain development and function are dependent on a fine-tuned balance of synaptic excitation and inhibition. As such, dysregulation of inhibition has been implicated in a variety of neurodevelopmental disorders. The cellular targets and consequences of GABAergic signaling in adult animals is well understood, however the specific interneurons modulated by ECB signaling remains poorly studied. Cortical inhibition is mediated by a diverse group of GABAergic interneurons (INs), including cells co-expressing the calcium binding protein parvalbumin (PV), the peptide transmitter somatostatin (SOM), or the serotonin 5HT3a receptor. Cortical INs differ in their synaptic targets, with PV-INs making synapses onto the perisomatic and proximal dendritic regions of target pyramidal neurons and SOM-INs making connections onto the dendritic arbors. A subset of 5HT3a-INs which express the vasoactive intestinal peptide (VIP), innervate dendrites of pyramidal neurons and other interneurons. This subcellular localization allows PV-INs to regulate the magnitude and timing of PN spike output and SOM-INs to regulate dendritic spine and shaft calcium influx along with glutamatergic synaptic plasticity. While the birth and migration of GABAergic INs is well understood, there remain gaps in our knowledge of the molecular and cellular mechanisms that influence inhibitory signaling. ECBs influence synaptogenesis and long-term plasticity of both excitatory and inhibitory connections. Although data has shown coexpression of the cannabinoid type-1 (CB1) receptor with SOM or VIP, colocalization of the CB1 receptor within the various interneurons has not been studied and will be addressed in this proposal. ECB release in the cortex modulates presynaptic GABA release and can drive both short- and long-term depression of inhibitory transmission, however the identity of the presynaptic INs sensitive to ECB activity is not well-characterized. Moreover, the consequences of CB1 receptor loss from distinct interneurons is unknown. In this proposal, we seek to determine how ECB signaling modulates GABAergic signaling mediated by distinct IN populations. We hypothesize that ECB signaling modulates cortical activity, with a key role in influencing plasticity mediated by dendritic targeting interneurons. We propose a novel combination of tools including electrophysiology, 2-photon laser-scanning microscopy (2PLSM), and optogenetic stimulation of genetically-targeted INs in the mouse primary visual cortex. Our experiments will provide an unprecedented level of insight into the development of GABAergic circuits in the neocortex.

该提案的总体目标是研究内源性大麻素 (ECB) 信号在 调节不同的皮质抑制性突触。大脑的生理发育和功能是 依赖于突触兴奋和抑制的微调平衡。因此，抑制失调 与多种神经发育障碍有关。细胞目标和后果 成年动物中的 GABA 信号传导已广为人知，然而，特定的中间神经元受 欧洲央行信号传导的研究仍然很少。皮质抑制是由多种 GABA 能介导的 中间神经元 (IN)，包括共表达钙结合蛋白小清蛋白 (PV) 的细胞，该肽 递质生长抑素 (SOM) 或血清素 5HT3a 受体。皮质 IN 的突触目标不同， PV-IN 在目标锥体神经元的体周和近端树突区域上形成突触 SOM-IN 连接到树突轴上。 5HT3a-IN 的一个子集，表达 血管活性肠肽（VIP），神经锥体神经元和其他中间神经元的树突。这 亚细胞定位允许 PV-IN 调节 PN 尖峰输出的幅度和时间，并且 SOM-IN 可以调节 调节树突棘和轴钙流入以及谷氨酸突触可塑性。虽然出生和 GABAergic IN 的迁移已被充分理解，但我们对分子和迁移的了解仍然存在差距。 影响抑制信号传导的细胞机制。 ECB 影响突触发生和长期可塑性 兴奋性和抑制性连接。 Although data has shown coexpression of the cannabinoid type-1 (CB1) 受体与 SOM 或 VIP，CB1 受体在各个中间神经元内的共定位没有 已被研究并将在本提案中得到解决。 ECB 在皮质中的释放调节突触前 GABA 释放并可以驱动抑制性传播的短期和长期抑制，但是 对 ECB 活动敏感的突触前 IN 尚未得到很好的表征。此外，CB1的后果 不同中间神经元的受体损失尚不清楚。在本提案中，我们寻求确定欧洲央行如何发出信号 调节由不同 IN 群体介导的 GABA 信号。我们假设欧洲央行信号 调节皮质活动，在影响树突靶向介导的可塑性方面发挥关键作用 中间神经元。我们提出了一种新颖的工具组合，包括电生理学、2 光子激光扫描 显微镜 (2PLSM) 和小鼠初级视觉中基因靶向 IN 的光遗传学刺激 皮质。我们的实验将为 GABAergic 的发展提供前所未有的洞察力 新皮质中的回路。