Photo-chemical tools for manipulating neural plasticity

描述（由申请人提供）：新蛋白质的合成与学习和记忆有根本联系。在分子水平上，有强有力的证据将特定类型的突触活动与新蛋白质的局部合成联系起来。这项拨款申请提出了创建新的光化学工具，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元细胞培养物、脑切片和最终在活体动物中产生的位置和时间。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组分的已知结构的化学合成。将产生众所周知的蛋白质合成抑制剂（嘌呤霉素、雷帕霉素、4 E结合蛋白）的光激活形式，包括基于偶氮苯光开关的可逆形式，所述偶氮苯光开关可以用光打开并且在黑暗中关闭。第二代光激活抑制剂的基础上的光活性黄蛋白/4 E结合蛋白融合的计划是遗传编码的。遗传编码的抑制剂可以在特定细胞中选择性地表达，从而能够在整个活体动物中对参与学习和记忆的分子事件进行明确的操纵。因此，这些工具的使用有望帮助揭示学习和记忆在分子水平上是如何在活体动物中发生的。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“错误布线”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有效治疗的途径。

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.