Function and Modulation of Somatic and Dendritic Kv3.3 Channels in Purkinje Cells

描述(由申请人提供)：离子通道中的点突变如何导致运动障碍、智力低下和大规模神经退化？最近发现钾通道Kv3.3突变是人类脊髓小脑性共济失调SCA13的原因。Kv3.3基因突变的患者表现为小脑萎缩，并出现严重的神经肌肉和认知症状。本研究旨在研究Kv3.3通道在小脑浦肯野细胞中的功能，为了解SCA13患者的疾病病理生理做出贡献。Kv3.3在Purkinje细胞中表达最强，蛋白定位于胞体、轴突和树突。浦肯野细胞的一种高度刻板印象的反应是复杂的棘波，这是一种对攀升纤维激活的大规模全有或全无反应，涉及体细胞和树突电活动以及大量树突钙离子瞬变。根据Kv3.3亚基的表达和电学性质，我们推测Kv3.3亚基在复杂的棘波中活跃，在调节树突状钙离子内流中起重要作用。我们打算用药理学和遗传学的方法来阐明体细胞和树突状Kv3.3通道在调节浦肯野细胞的电特性和钙动态中的特定功能。钙稳态的改变可能是Kv3.3突变人类的病理原因之一，这些研究对于确定这些通道在调节钙动态中的作用是至关重要的。我们实验室的研究证实了在异源表达系统中，PKC对Kv3.3亚基的调控。PKC在浦肯野细胞的功能和可塑性中起着非常重要的作用，Kv3.3的这种调节可能是PKC细胞效应的一个重要机制。此外，PKCγ基因突变是人类脊髓小脑性共济失调SCA14的原因。因此，Kv3.3或PKC Gamma的突变产生相似的表型，提出了一种有趣的可能性，即Kv3.3通道的调制参与了PKC Gamma突变的人类疾病的表达。这项研究的第二个目的是证明浦肯野细胞中Kv3.3亚单位的调节，并开始探索这种调节对浦肯野细胞生理学的影响。

DESCRIPTION (provided by applicant): How can a point mutation in an ion channel cause motor deficits, mental retardation and massive neurodegeneration? It was recently discovered that mutations in potassium channel Kv3.3 is the cause of human spinocerebellar ataxia SCA13. Affected individuals with mutations in Kv3.3 display cerebellar atrophy and present with severe neuromuscular and cognitive symptoms. This research focuses on the function of Kv3.3 channels in Purkinje cells of the cerebellum, and is a contribution in understanding the pathophysiology of disease in humans with SCA13. Kv3.3 is most strongly expressed in Purkinje cells, with protein localization in somas, axons and dendrites. A highly stereotyped response of Purkinje cells is the complex spike, a massive all-or-none response to climbing fiber activation involving somatic and dendritic electrical activity and large dendritic Ca ++ transients. Due to the expression and electrical properties of Kv3.3 subunits, we hypothesize that they are active in the complex spike and important in the regulation dendritic Ca++ influx. We intend to use pharmacological and genetic approaches to elucidate the specific functions of somatic and dendritic Kv3.3 channels in modulating the electrical properties and Ca++ dynamics of Purkinje cells. Altered Ca++ homeostasis is a probable cause of pathology in humans with Kv3.3 mutations, and these studies are critical in establishing the roles of these channels in regulating Ca++ dynamics. Studies in our laboratory demonstrated the modulation of Kv3.3 subunits by PKC in heterologous expression systems. PKC plays very important roles in Purkinje cell function and plasticity, and it is possible that this modulation of Kv3.3 is an important mechanism in the cellular effects of PKC. Moreover, mutations in PKC gamma are the cause of human spinocerebellar ataxia SCA14. Thus, mutations in either Kv3.3 or PKC gamma produce similar phenotypes, posing the intriguing possibility that modulation of Kv3.3 channels is involved in the expression of disease in humans with mutations in PKC gamma. A second aim of this study is to demonstrate the modulation and Kv3.3 subunits in Purkinje cells and begin to explore the implications of this modulation on Purkinje cell physiology.