Photo-chemical tools for manipulating neural plasticity

描述(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，有强有力的证据表明特定类型的突触活动与局部合成新蛋白质有关。这项拨款申请建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，从而控制在神经细胞培养中、在脑片中以及最终在活动物中制造蛋白质的地点和时间。创建这些工具所采用的方法将是基于产生新蛋白质的生化途径的关键成分的已知结构的化学合成。将创造出众所周知的蛋白质合成抑制剂(嘌罗霉素、雷帕霉素、4E结合蛋白)的光激活版本，包括基于偶氮苯光开关的可逆版本，这种开关可以用光打开，也可以在黑暗中关闭。基于光活性黄色蛋白/4E结合蛋白融合的第二代光激活抑制剂正在计划中，它是遗传编码的。基因编码的抑制物可以在特定的细胞中选择性地表达，从而能够对整个活着的动物学习和记忆中涉及的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着的动物是如何学习和记忆的。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。在分子水平上了解这是如何起作用的，将为有效治疗指明道路。

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.