Identification of the Protein Kinases which Regulate Mammalian Synapse Formation

描述（由申请人提供）：突触是神经系统中细胞与细胞接触的特殊部位，介导神经元之间的交流。哺乳动物中枢神经系统中这些关键结构形成的分子机制仍有待发现。在此之前，我们开发了一种基于rna干扰（RNAi）的新型正向遗传方法来识别调节突触形成的新分子。目前，我们正在应用这项技术来了解药物成瘾的细胞生物学。到目前为止，已经有许多激酶参与调节突触的形成或功能，这为蛋白激酶在突触中具有关键功能的假设提供了支持。此外，蛋白激酶信号的激活被假设为药物暴露后神经元结构和突触变化的基础。因此，进一步研究蛋白激酶在突触形成中的作用是必要的。为此，我们建议采用全基因组方法来鉴定在培养的哺乳动物神经元突触形成时表达的蛋白激酶的完整补体。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能和/或gaba能突触所必需的。公共卫生相关性：目前有一种假说解释了药物成瘾的持续特征，包括药物渴望和复发，该假说认为突触结构和神经元连通性的变化是该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，一种全基因组的方法来理解蛋白激酶在突触形成和功能中的作用，如本提案所述，有可能对药物成瘾的一部分特征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.