The Essential Role of Presynaptic NMDA Receptors in The Fast Antidepressant Actions of Ketamine and Its Metabolite

突触前 NMDA 受体在氯胺酮及其代谢物快速抗抑郁作用中的重要作用

• 批准号: 9552924
• 负责人: Xiang Cai
• 金额: $ 33.19万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9552924
• 关键词: AMPA Receptors; Adverse effects; Adverse reactions; Animal Experiments; Antidepressive Agents; Behavior; Behavioral; Biological; Biotinylation; Cell surface; Cells; Chemosensitization; Clinical Trials; Development; Dialysis procedure; Dose; Down-Regulation; Electrophysiology (science); Evaluation; Event; Exhibits; Exocytosis; Experimental Designs; GABA Receptor; Generations; Glutamate Receptor; Glutamates; HCN1 gene; Hippocampus (Brain); In Vitro; Ketamine; Knock-in Mouse; Knockout Mice; Knowledge; Location; MK801; Measurement; Mediating; Membrane Proteins; Mental Depression; Molecular; N-Methyl-D-Aspartate Receptors; NR1 gene; Neurons; PKA inhibitor; Patients; Phosphorylation; Property; Proteins; Pyramidal Cells; Receptor Inhibition; Reporting; Role; Signal Transduction; Site; Surface; Synapses; Synaptic Transmission; Techniques; Testing; Therapeutic; Western Blotting; antidepressant effect; base; behavior measurement; club drug; depressive symptoms; design; experimental study; hippocampal pyramidal neuron; in vivo; multidisciplinary; next generation; patch clamp; postsynaptic; presynaptic; prevent; response; treatment-resistant depression

Project Summary/Abstract Evidence from clinical trials and animal experiments shows a single subanesthesia dose of ketamine produces rapid antidepressant responses. However, the psychotomimetic properties and abuse potential of ketamine necessitate caution in promoting this compound to be used as a general treatment of depression. The reports on the mechanisms underlying the rapid-acting antidepressant effects of ketamine are controversial. Particularly, it remains uncertain which molecular events mediate ketamine’s or its metabolites’ action in enhancing excitatory synaptic transmission. In preliminary experiments, it was observed that both ketamine and its metabolite, cis-6-HNK, potently enhanced Shaffer collateral-CA1 (SC-CA1) EPSCs and fEPSPs. These actions were mimicked and completely occluded by MK-801, but not eliminated by GABA receptor blockade. Ketamine potently increased total and surface GluA1 expression and the phosphorylation of GluA1 Ser845 site. In addition, ketamine failed to potentiate SC-CA1 fEPSPs and induce the antidepressant-like responses in GluA1 S845A knock-in mice. Furthermore, deletion of NMDA receptor on CA3 neurons but not on CA1 cells completely eliminated ketamine-induced potentiation on SC-CA1 synaptic transmission. In the proposed project experiments are designed to test the role of GluA1 Ser845 phosphorylation and presynaptic NMDA receptors in ketamine- and HNK-produced rapid-acting antidepressant responses as well as their celluar and molecular bases. Electrophysiological techniques will be combined, including whole-cell patch clamp with cell biological technique such as Western blotting, biotinylation of surface proteins and GluR1 S845A knock-in mice, CA1 cell- or CA3 cell-specific NR1 knockout mice, HCN1 knockout mice as well as behavioral measurements to achieve the designed experiments. The proposed project will expand knowledge in understanding the cellular mechanisms of fast-acting antidepressant actions of ketamine. A better understanding of ketamine’s fast-acting antidepressant effects will promote the design of a new generation of rapid-acting antidepressant to offer a better therapy to depression.

项目概要/摘要 临床试验和动物实验的证据表明，单次亚麻醉剂量 氯胺酮可产生快速的抗抑郁反应，但具有拟精神病作用。 和氯胺酮滥用的可能性，因此在推广该化合物作为药物时必须谨慎 关于快速作用机制的报告。 氯胺酮的抗抑郁作用目前尚不确定。 分子事件介导氯胺酮或其代谢物增强兴奋性突触的作用 在初步实验中，观察到氯胺酮及其代谢物， cis-6-HNK，有效增强 Shaffer 侧支 CA1 (SC-CA1) EPSC 和 fEPSP。 动作被 MK-801 模仿并完全遮挡，但未被 GABA 消除 受体阻断。氯胺酮有效增加总 GluA1 表达和表面 GluA1 表达。 GluA1 Ser845 位点的磷酸化此外，氯胺酮未能增强 SC-CA1 fEPSP。 并在 GluA1 S845A 敲入小鼠中诱导抗抑郁样反应。此外， CA3 神经元上的 NMDA 受体缺失，但 CA1 细胞上的 NMDA 受体缺失完全消除 在拟议的项目中，氯胺酮诱导的 SC-CA1 突触传递增强。 实验旨在测试 GluA1 Ser845 磷酸化和突触前的作用 氯胺酮和 HNK 中的 NMDA 受体产生快速抗抑郁反应 以及他们的细胞和分子基础将被结合起来， 包括采用蛋白质印迹等细胞生物学技术的全细胞膜片钳， 表面蛋白和 GluR1 S845A 敲入小鼠的生物素化，CA1 细胞或 CA3 细胞特异性 NR1 敲除小鼠、HCN1 敲除小鼠以及行为测量以实现 所设计的实验将扩展对理解的知识。 更好地了解氯胺酮快速抗抑郁作用的细胞机制。 氯胺酮快速抗抑郁作用的研究将促进新一代药物的设计 速效抗抑郁药，为抑郁症提供更好的治疗方法。