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: 7688279
• 负责人: Andrew Woolley
• 金额: $ 17.06万
• 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/7688279
• 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; Learning; Life; Light; Link; Memory; Mental Health; 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; public health relevance; 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（由申请人提供）：新蛋白质的合成从根本上与学习和记忆有关。在分子水平上，有强有力的证据将特定类型的突触活性与新蛋白质的局部合成联系起来。该赠款的应用提出了创建新的光化学工具，该工具将使研究人员能够使用光来操纵翻译，从而控制神经元细胞培养物，脑切片和最终在活的动物中制造蛋白质的位置和何时。创建这些工具的方法将基于生产新蛋白质的生化途径的关键组成部分的已知结构化学合成。将创建众所周知的蛋白质合成抑制剂的光活化版本（嘌呤霉素，雷帕霉素，4E结合蛋白），包括基于偶氮苯光相关的可逆版本，可以用光打开并在黑暗中关闭。计划了基于光活性黄色蛋白/4E结合蛋白融合的第二代光活化抑制剂，该抑制剂已计划在遗传编码中。可以在特定细胞中选择性地表达遗传编码的抑制剂，从而实现对整个活动物中学习和记忆涉及的分子事件的明确定义的操纵。因此，这些工具的使用有望帮助揭示生物水平的生物中的学习和记忆如何发生。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，强迫性疾病或成瘾，与大脑的“误会”有关，即异常的突触可塑性有关。了解这在分子水平上的工作方式将显示有效治疗的途径。