Role of KCTD proteins in striatal signaling

项目摘要 纹状体cAMP信号转导的改变有助于运动障碍的进展，其 不断增加的流行率是一个重大的公共卫生问题。运动障碍的治疗选择 部分受限于对纹状体神经元中cAMP信号的分子控制的了解不足。 尽管在解开cAMP通路方面取得了巨大的进展，但cAMP的复杂性 纹状体的调节，加上大量发挥关键作用的球员尚未完全阐明 减缓了朝着更有效的护理战略的进展。为了克服这一障碍，我们的长期目标是 更好地理解控制纹状体cAMP信号的机制和原理。拟议中的工作 主要介绍钾通道四聚化(KCTD)蛋白在纹状体信号调节中的作用 通过G蛋白偶联受体(GPCRs)。KCTD蛋白最近参与了gpr信号转导。 直接通过与G蛋白结合，或通过锌转运蛋白间接影响锌的内流。 然而，KCTD调节G蛋白信号的细节以及锌对纹状体信号的影响 才刚刚开始出现。这项建议的目的是阐明KCTD-锌-cAMP信号轴在 天然的神经环境。在目标1中，我们将使用生化分析来研究负反馈。 动态调节纹状体KCTD表达的压力，然后通过光学分析来研究如何 KCTD调节G蛋白与效应物的相互作用。AIM 2将利用生化分析和光学技术 活体神经元中的传感器，以描绘KCTD和锌在纹状体cAMP信号上的精确机制细节。 在目标3中，我们将讨论KCTD如何通过光刺激来塑造本地生理信号询问 神经调节性释放与信号整合的并行双光子成像 表达光学感受器的基因定义的纹状体神经元。总的来说，拟议的研究预计将 重要的是，它将提供对KCTD-锌-cAMP的基本机制和生理洞察 对运动生理学至关重要的信号传递过程的影响。

Project Summary Alterations in striatal cAMP signal transduction contribute to the progression of movement disorders, whose increasing prevalence present a significant public health concern. Therapeutic options for movement disorders are limited in part by a deficient understanding of the molecular control of cAMP signaling in striatal neurons. Although tremendous progress has been made in unraveling the cAMP pathway, complexity of cAMP regulation in the striatum coupled with the large number of players with pivotal roles not yet fully elucidated has slowed progress towards more effective care strategies. To overcome this barrier, our long-term goal is to better understand the mechanisms and principles that control striatal cAMP signaling. The proposed work focuses on the role of Potassium Channel Tetramerization (KCTD) proteins in modulation of striatal signaling through G protein coupled receptors (GPCRs). KCTD proteins were recently implicated in GPCR signaling both directly by engaging with G proteins and indirectly by influencing zinc influx through zinc transporters. However, the details of KCTD regulation of G protein signaling and consequences of zinc on striatal signaling are only beginning to emerge. The goal of this proposal is elucidation of the KCTD-Zinc-cAMP signaling axis in the native neuronal environment. In Aim 1, we will use biochemical assays to study negative feedback pressure that dynamically adjusts KCTD expression in the striatum followed by optical assays to study how KCTD regulate interaction of G proteins with effectors. Aim 2 will utilize biochemical assays and optical sensors in live neurons to delineate precise mechanistic details of KCTD and zinc on striatal cAMP signaling. In Aim 3, we will address how KCTDs shape native physiological signal interrogation by optically stimulating neuromodulatory release with concurrent 2-photon imaging of signal integration across intact circuits in genetically defined striatal neurons expressing optical sensors. In total, the proposed research is expected to be significant in that it will provide essential mechanistic and physiological insight into the KCTD-Zinc-cAMP impact on signaling processes critical for motor physiology.