Photo-chemical tools for manipulating neural plasticity

用于操纵神经可塑性的光化学工具

• 批准号: 8101113
• 负责人: Andrew Woolley
• 金额: $ 16.38万
• 依托单位: UNIVERSITY OF TORONTO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8101113
• 关键词: Animals; Applications Grants; Binding; Binding Proteins; Biochemical Pathway; Brain; Cell Culture Techniques; Cells; Chemicals; Cordycepin; Data; Enzymes; Epilepsy; Event; Generations; Genetic Techniques; Goals; Health; Learning; Life; Light; Link; Memory; Mental Health; Messenger RNA; Molecular; Molecular Genetics; Nervous system structure; Neuronal Plasticity; Neurons; Obsessive-Compulsive Disorder; Pathway interactions; Peptides; Phosphotransferases; Poly A; Polyadenylation; Post-Traumatic Stress Disorders; Process; Protein Biosynthesis; Protein Synthesis Inhibitors; Proteins; Puromycin; Regulation; Research Personnel; Ribosomes; Role; Signal Transduction; Signal Transduction Pathway; Sirolimus; Slice; Structure; Synapses; Synaptic plasticity; Translation Initiation; Translations; Work; addiction; azobenzene; base; chemical synthesis; design; effective therapy; human FRAP1 protein; in vivo; inhibitor/antagonist; irradiation; mTOR Inhibitor; mTOR inhibition; neural circuit; penetratin; photo switch; polypeptide; prevent; protein aminoacid sequence; response; selective expression; tool; uptake

DESCRIPTION (provided by applicant): The synthesis of new proteins is fundamentally linked with learning and memory. At the molecular level there is strong evidence linking specific types of synaptic activity with the local synthesis of new protein. This grant application proposes the creation of new photochemical tools that will enable researchers to use light to manipulate translation and thereby control where and when proteins are made in neuronal cell culture, in brain slices and ultimately in living animals. The approach taken to the creation of these tools will be chemical synthesis based on the known structures of key components of the biochemical pathways that produce new proteins. Light- activated versions of well-known protein synthesis inhibitors (puromycin, rapamycin, 4E binding proteins) will be created, including reversible versions based on azobenzene photo-switches that can be turned on with light and that turn off in the dark. A second generation of light-activated inhibitors based on photo-active yellow protein/4E binding protein fusions is planned that is genetically-encoded. Genetically-encoded inhibitors can be selectively expressed in particular cells thereby enabling well-defined manipulation of the molecular events involved in learning and memory in whole living animals. Use of these tools thus promises to help uncover how learning and memory occur in living animals at a molecular level. PUBLIC HEALTH RELEVANCE: Numerous mental health problems including post-traumatic stress disorder, epilepsy, obsessive compulsive disorders, or addiction are connected with "mis-wiring" of the brain, i.e. aberrant synaptic plasticity. Understanding how this works at the molecular level will show the path to effective treatment.

描述（申请人提供）：新蛋白质的合成与学习和记忆有着根本的联系。在分子水平上，有强有力的证据表明，特定类型的突触活动与新蛋白质的局部合成有关。这项拨款申请提议创造新的光化学工具，使研究人员能够利用光来操纵翻译，从而控制蛋白质在神经细胞培养、大脑切片以及最终在活体动物中的制造地点和时间。创造这些工具的方法将是基于产生新蛋白质的生化途径关键成分的已知结构的化学合成。众所周知的蛋白质合成抑制剂（嘌呤霉素、雷帕霉素、4E结合蛋白）的光激活版本将被创造出来，包括基于偶氮苯光开关的可逆版本，它可以在光照下打开，在黑暗中关闭。基于光活性黄蛋白/4E结合蛋白融合体的第二代光激活抑制剂计划被遗传编码。基因编码的抑制剂可以选择性地表达在特定的细胞中，从而能够在整个活体动物的学习和记忆过程中对分子事件进行明确的操作。因此，使用这些工具有望在分子水平上帮助揭示活体动物的学习和记忆是如何发生的。公共卫生相关性：许多心理健康问题，包括创伤后应激障碍、癫痫、强迫症或成瘾，都与大脑的“错误连接”有关，即异常的突触可塑性。了解这在分子水平上是如何起作用的，将显示出有效治疗的途径。