Cellular regulation of Sodium-activated Ion Channels

描述（由申请人提供）：Slack和Slick基因编码Na+激活的K+通道，其调节神经元适应维持突触刺激的速度和神经元动作电位计时的准确性。这些通道也被认为在保护神经元和心肌细胞免受缺氧损伤中起关键作用。在它们的总体结构上，它们类似于其他电压门控K通道，但具有非常大的（bbb600个氨基酸）细胞内c端。Slack的c端结构域与脆性- X智力迟钝蛋白（FMRP）相互作用，FMRP是一种rna结合蛋白，调节神经元mrna子集的运输和翻译。本应用程序的工作将使用生化和电生理分析来确定Slack和FMRP在体外和体内相互作用中的特定区域，并确定FMRP需要哪些特定区域来控制Slack通道的门控。我们还将确定在脆性X综合征（Fmr1-/-）小鼠模型中Na+激活的K+通道的表达是如何改变的。特别是，我们将确定在Fmr1-/-动物中导致Slack远端C-末端表达完全丧失的机制，该区域似乎是FMRP与通道结合所必需的。平行的电生理和药理学实验将评估Slack表达模式的改变对天然神经元中Na+激活的K+通道功能特性的影响。了解Slack和Slick通道的生物学特性和调控将有助于更清楚地了解脆性X综合征和相关疾病的缺陷，并有望导致新的治疗策略的发展。

DESCRIPTION (provided by applicant): The Slack and Slick genes encode Na+-activated K+ channels, which regulate the rate at which neurons adapt to maintained synaptic stimulation and the accuracy of timing of neuronal action potentials. These channels have also been proposed to play a key role in the protection of neurons and cardiomyocytes from hypoxic injury. In their general structure, they resemble other voltage-gated K channels, but have very large (>600 amino acid) intracellular C-termini. The C-terminal domain of Slack interacts with Fragile- X Mental Retardation protein (FMRP), an RNA-binding protein that regulates trafficking and translation of a subset of subset of neuronal mRNAs. The work in this application will use biochemical and electrophysiological assays to determine the specific regions of Slack and FMRP involved in their interactions both in vitro and in vivo, and will determine which specific regions are required for FMRP to control the gating of Slack channels. We shall also determine how the expression of Na+-activated K+ channels is altered in a mouse model of Fragile X syndrome (Fmr1-/-). In particular we shall determine the mechanism that in Fmr1-/- animals causes the total loss of expression of the distal C- terminus of Slack, a region that appears to be required for FMRP binding to the channels. Parallel electrophysiological and pharmacological experiments will evaluate the effects of this altered pattern of Slack expression on the functional properties of Na+- activated K+ channels in native neurons. An understanding of the biological properties and regulation of Slack and Slick channels will lead to a clearer understanding of the deficits in Fragile X syndrome and related disorders and is expected to lead to the development of novel therapeutic strategies.