Neuronal Gαi-GPCR targeting to primary cilia and impact on cAMP mediated transcription

项目总结/摘要 纤毛病是一组疾病，其特征是基因突变损害初级纤毛形态或 功能，并以包括发育障碍、肥胖、认知和 人类的行为缺陷初级纤毛是从神经元索马体伸出的不动细胞器， 靠近核室。越来越多的证据表明，细胞外刺激（即， 神经调质）通过位于神经元上的G蛋白偶联受体（GPCR）传递专门的信号传导。 神经元初级纤毛影响转录变化和神经元活性。因此，确定如何 特异性靶向神经元睫状GPCR，将导致神经和 神经精神疾病该提案将侧重于确定所采用的选择性纤毛靶向机制 通过神经Gα i偶联GPCR和纤毛GPCR信号传导对基因转录的影响。 我们发现，在脑切片中，神经Gα i偶联的GPCR，如μ阿片受体，表达在脑组织中。 一些神经元群体的神经元初级纤毛，但其他人没有，表明细胞和/或受体选择性 纤毛靶向机制。在特定目标1中，我们分析了纤毛靶向机制是否是细胞特异性的 和/或在三种不同的细胞类型中具有受体选择性。在具体目标2中，我们采用基于蛋白质组学的方法 鉴定参与细胞和脑中Gα i偶联GPCR纤毛靶向的新型GPCR相关蛋白 组织.在具体目标3中，我们将评估纤毛Gα i偶联的GPCR环AMP信号转导对 神经元转录总的来说，这项建议将试图辨别潜在的机制纤毛 靶向神经元GPCR和睫状GPCR的神经调节功能。

PROJECT SUMMARY / ABSTRACT Ciliopathies are a collection of diseases that feature genetic mutations that impair primary cilia morphology or function and are characterized by symptoms that include developmental disorders, obesity, cognitive and behavioral deficits in humans. Primary cilia are immotile organelles that protrude off neuronal soma often adjacent to the nuclear compartment. A growing body of evidence indicates that extracellular stimuli (i.e., neuromodulators) transmits specialized signaling through G protein-coupled receptors (GPCRs) located on neuronal primary cilia to influence transcriptional changes and neuronal activity. Thus, determining how to specifically target neuronal ciliary GPCRs, will lead to novel therapeutic approaches for neurological and neuropsychiatric diseases. This proposal will focus on identifying selective ciliary targeting mechanics employed by neural Gαi-coupled GPCRs and the impact of ciliary GPCR signaling on gene transcription. We found that in brain sections, neural Gαi-coupled GPCRs, such as the mu opioid receptor, are expressed in neuronal primary cilia of some neuronal populations but not others suggesting cell and/or receptor-selective ciliary targeting machinery. In Specific Aim 1, we dissect whether ciliary targeting mechanics are cell-specific and/or receptor-selective in three distinct cell types. In Specific Aim 2, we employ Proteomics-based approaches to identify novel GPCR-associated proteins involved in ciliary targeting of Gαi-coupled GPCRs in cells and brain tissue. In Specific Aim 3, we will evaluate the contribution of ciliary Gαi-coupled GPCR cyclic-AMP signaling to neuronal transcription. Collectively, this proposal will attempt to discern the mechanisms underlying ciliary targeting of neuronal GPCRs and the neuromodulatory function of ciliary GPCRs.