Identification of the Protein Kinases which Regulate Mammalian Synapse Formation

描述(申请人提供)：突触是细胞-细胞接触的专门位置，在神经系统中的神经元之间进行沟通。关于哺乳动物中枢神经系统中这些关键结构形成的分子机制，还有很多有待发现的地方。在此之前，我们开发了一种基于正向遗传RNA干扰(RNAi)的新方法来识别调节突触形成的新分子。目前，我们正在将这项技术应用于了解药物成瘾的细胞生物学。到目前为止，已有许多蛋白激酶参与调节突触的形成或功能，这支持了蛋白激酶在突触中具有关键功能的假说。此外，蛋白激酶信号的激活被认为是应对药物暴露而导致神经元结构和突触变化的基础。因此，有必要进一步研究蛋白激酶在突触形成中的作用。为此，我们建议采用全基因组的方法来确定在培养的哺乳动物神经元中形成突触时表达的蛋白激酶的全部补充。接下来，我们将利用我们基于RNAi的筛选方法来询问哪些激酶是形成功能性谷氨酸和/或GABA能突触所必需的。与公共健康相关：目前一种解释药物成瘾持续特征的假说，包括对药物的渴望和复发，假设突触结构和神经元连接的变化是疾病这些特征的基础。此外，蛋白激酶的功能与突触结构的这些变化有关。因此，本提案中概述的了解蛋白激酶在突触形成和功能中的作用的全基因组方法有可能对药物成瘾特征的子集的潜在原因产生重要的见解。

DESCRIPTION (provided by applicant): Synapses are specialized sites of cell-cell contact that mediate communication between neurons in the nervous system. Much remains to be discovered about the molecular mechanisms that underlie formation of these critical structures in the mammalian central nervous system. Previously, we developed a novel, forward genetic, RNA-interference (RNAi)-based approach to identifying new molecules that regulate synapse formation. Currently, we are in the process of applying this technology to understanding the cell biology of drug addiction. Thus far, a number of kinases have been implicated in regulating synapse formation or function, lending support to the hypothesis that protein kinases have critical functions at the synapse. Further, activation of protein kinase signaling has been hypothesized to underlie changes in neuronal structure and synapses in response to drug exposure. Therefore, further investigation into the role of protein kinases in synapse formation is warranted. To this end, we propose to take a genome-wide approach to identify the full complement of protein kinases that are expressed at the time that synapses are forming in cultured mammalian neurons. Next, we will utilize our RNAi-based screening approach to ask which kinases are required for the formation of functional glutamatergic and/or GABAergic synapses. PUBLIC HEALTH RELEVANCE: A current hypothesis to explain the persistent features of drug addiction, including drug cravings and relapse, posits that changes in synaptic structure and neuronal connectivity underlie these features of the disease. Further, the function of protein kinases has been implicated in these changes in synaptic structure. Thus, a genome-wide approach to understanding the role of protein kinases in synapse formation and function as outlined in this proposal has the potential to yield important insights into the underlying causes of a subset of features of drug addiction.