Molecular Properties of Neuronal Dendritic K+ Channels

我们的一般工作假设是，天然电压门控钾通道的功能特性是通过不同辅助蛋白与核心孔形成α亚基的相互作用而产生的。这一假说强调了蛋白质-蛋白质相互作用在神经元电特性的形成、运输和调节中的重要性。在这个方案中，我们将检验Kv4α亚基，也称为Shal型Kv通道，与辅助KChlP蛋白和DPP蛋白组装形成大分子蛋白复合体的特定假设，该复合体被输送到CA1锥体神经元的树突，形成功能上重要的A电流通道。我们的研究将集中于分析天然神经元A电流功能的变化，以响应分子遗传操作的表达水平 天然神经元中的不同亚基。接下来，我们将研究引入重要亚基蛋白质的突变版本的功能影响，这些蛋白质要么缺乏执行特定蛋白质-蛋白质相互作用的能力，要么无法被特定的翻译后修饰所修饰。通过描述不同水平上的功能变化，我们将确定不同的亚单位蛋白、功能基序和翻译后修饰在形成CA1锥体神经元A电流表型中的具体作用。这些研究将对我们理解神经元电表型的分子控制以及理解其他离子通道和辅助亚单位蛋白的功能重要性的方法产生普遍的影响。我们的研究将对 我们对兴奋性调节的理解将影响许多健康相关研究领域，包括癫痫、心脏病、哮喘和艾兹海默氏症。

Our general working hypothesis is that the functional properties of native voltage-gated potassium channels arise through the interactions of different auxiliary proteins with the core pore forming alpha subunits. This hypothesis emphasizes the importance of protein-protein interactions in the formation, trafficking, and regulation of neuronal electrical properties. In this proposal we will be testing the specific hypothesis that Kv4 alpha subunits, also called Shal type Kv channels, assemble with auxiliary KChlP proteins and DPP proteins to form a macromolecular protein complex that is trafficked to the dendrites in CA1 pyramidal neurons to form a functionally important A current channel. Our studies will focus on the analysis of changes in A current function in native neurons in response to molecular genetic manipulations in expression level for different subunits in native neurons. Next, we will examine the functional effects of introducing mutant version of the important subunits proteins that either lack the ability to perform specific protein-protein interactions or are unable to be modified by particular post-translational modifications. By characterizing the changes in function at several different levels, we will determine the specific roles of different subunit proteins, functional motifs, and post-translational modifications in forming the A current phenotypes of CA1 pyramidal neurons. These studies will have a general impact on our understanding of the molecular control of neuronal electrical phenotypes as well as the approaches we will take in understand the functional importance of other ion channels and auxiliary subunit proteins. Our studies will have a general impact on our understanding of the regulation of excitability that will impact many areas of health related research including epilepsy, heart disease, asthma, and AIzheimer's.