Cellular regulation of Sodium-activated Ion Channels

钠激活离子通道的细胞调节

基本信息

批准号：
8337337
负责人：
LEONARD K KACZMAREK
金额：
$ 35.17万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-26 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8337337
关键词：
Action Potentials Adult Affect Amino Acid Sequence Amino Acids Animals Antibodies Architecture Binding Biochemical Biological Biological Assay Brain C-terminal Cardiac Myocytes Cell membrane Cells Chemosensitization Development Disease Distal Drug Delivery Systems Epitopes FMR1 Face Fluorescence Resonance Energy Transfer Fragile X Mental Retardation Protein Fragile X Syndrome Genes Genetic Human Hypoxia Immunoprecipitation Impairment In Vitro Inherited Injury Intellectual functioning disability Ion Channel Knockout Mice Lead Mass Spectrum Analysis Membrane Proteins Messenger RNA Molecular Conformation Mus Mutate N-terminal Neurodevelopmental Disorder Neurons Pattern Peptides Play Potassium Channel Property Protein Binding Protein Binding Domain Protein Isoforms Proteins RNA Binding RNA Splicing RNA-Binding Proteins Regulation Role Site-Directed Mutagenesis Slice Sodium Structure Synapses Testing Time Translations Two-Hybrid System Techniques Voltage-Gated Potassium Channel Work Yeasts cell growth regulation immunoreactivity in vivo mouse model mutant novel therapeutics protein function protein protein interaction research study trafficking

项目摘要

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.

描述（由申请人提供）：松弛和光滑基因编码Na+激活的K+通道，该通道调节神经元适应维持的突触刺激的速率和神经元动作电位的准确性。还提出了这些通道在保护神经元和心肌细胞免受低氧损伤的保护中发挥关键作用。在其一般结构中，它们类似于其他电压门控的K通道，但具有非常大的（> 600个氨基酸）内C-termini。 Slack的C末端结构域与脆弱的X智力低下蛋白（FMRP）相互作用，这是一种RNA结合蛋白，可调节贩运和翻译神经元mRNA的子集的子集。本应用中的工作将使用生化和电生理测定法来确定其在体外和体内相互作用涉及的松弛和FMRP的特定区域，并将确定FMRP需要哪些特定区域来控制松弛通道的门控。我们还将确定在脆弱X综合征的小鼠模型（FMR1 - / - ）中如何改变Na+活化的K+通道的表达。特别是，我们将确定在FMR1 - / - 动物中导致Slack远端C末端的总表达损失的机制，Slack的远端C末端是FMRP与通道结合所必需的区域。平行的电生理和药理实验将评估这种松弛表达模式对天然神经元中Na+活化的K+通道功能特性的改变的影响。对松弛和光滑通道的生物学特性和调节的理解将使人们对脆弱X综合征和相关疾病的缺陷有更清晰的了解，并有望导致新的治疗策略的发展。