Mechanism and functional role of AKAP79/150 in M current control

AKAP79/150 在 M 电流控制中的机制和功能作用

基本信息

批准号：
8549448
负责人：
MARK S SHAPIRO
金额：
$ 46.5万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2014-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8549448
关键词：
A kinase anchoring protein Acetylcholine Afferent Neurons Agonist Angiotensin II Animals Binding Binding Sites Biological Biological Assay Bradykinin Brain Brain Stem Calcineurin Calmodulin Capsaicin Cell Communication Cell Nucleus Cell physiology Cells Confocal Microscopy Coughing Cyclic AMP-Dependent Protein Kinases Cyclosporine Cytoplasm DNA Dependence Depressed mood Dimerization Disease Dose Egtazic Acid Electrophysiology (science)Elements Emotional Enhancers Esthesia Family Feedback Fura-2 G Protein-Coupled Receptor Genes G-Protein-Coupled Receptors Gene Expression Genes Genetic Transcription Genomics Health Hippocampus (Brain)Histamine Image Inflammation Ion Channel Irritants Life Light Link Luc Gene Luciferases Measures Mediating Membrane Memory Messenger RNA Methods Modeling Molecular Moods Mus Mutation Nervous System Physiology Nervous system structure Neurons Nodose Ganglion Organ PC12 Cells Pain Pain Measurement Peripheral Personality Pharmaceutical Preparations Physiological Play Plethysmography Potassium Channel Protein Dephosphorylation Protein Isoforms Protein Kinase C Purinoceptor Rattus Receptor Signaling Recruitment Activity Regulation Regulatory Element Reporter Reverse Transcriptase Polymerase Chain Reaction Rodent Role Seizures Sensory Shapes Signal Pathway Signal Transduction Site Structure of superior cervical ganglion T-Cell Activation Techniques Testing Time Transcriptional Regulation Up-Regulation VIVIT peptide Visceral Visceral Afferents cellular imaging channel blockers chromatin immunoprecipitation constriction ganglion cell knock-down member mouse model mutant neuronal excitability nuclear factors of activated T-cells patch clamp promoter receptor receptor coupling research study response sensor small hairpin RNA transcription factor voltage

项目摘要

DESCRIPTION (provided by applicant): A-kinase anchoring proteins (AKAPs) organize numerous intracellular signaling pathways by bringing together their molecular components into discrete sub-cellular microdomains. One such AKAP, AKAP79/150 interacts with protein kinase A, protein kinase C (PKC), calmodulin (CaM), calcineurin (CaN), ion channels such as M-type (KCNQ, Kv7) K+ channels, L-type Ca2+ channels and G protein-coupled receptors. The nuclear factor of activated T-cells (NFAT) family of transcription factors has been shown to underlie activity-dependent regulation of gene transcription in excitable cells, including central and peripheral neurons. Sensory neurons of the nodose ganglia are responsible for sending afferents from the visceral organs to the brainstem, where body organs are controlled. In this project, we will study the transcriptional regulation of KCNQ2-3 channels in superior cervical ganglion (SCG) sympathetic neurons via NFATc1-c4 members, which are activated by intracellular Ca2+ (Ca2+i) signals, such as those generated by activity-dependent depolarization. We hypothesize the Ca2+i signals are sensed by CaN, anchored by AKAP79/150 to L-type CaV1.3 Ca2+ channels, followed by dephosphorylation of NFATs, their import into the nucleus, and up-regulation of M-channel gene expression that acts as a negative feedback on excitability in physiological and pathophysiological states. We will test this hypothesis using a variety of cutting-edge approaches. We further hypothesize AKAP79/150 to orchestrate modulation of M current in NG neurons by a variety of Gq/11-linked receptor types that are critical to pain and inflammation, including bradykinin, PAR-2, histamine, angiotensin II and purinergic receptors. We will determine which Gq/11-coupled receptors modulate NG neurons, and the role of AKAP79/150 in coupling the receptors to the M-type channels. Techniques to be used include patch-clamp electrophysiology of native SCG and NG neurons, single live-cell imaging, confocal microscopy, luciferase gene-reporter assays, genetically-altered mice and live-animal airway plethysmography. We aspire to discover the mechanisms endowing AKAP79/150 in organizing the transcriptional regulation of M-type K+ channels, and in orchestrating the receptor signaling towards these channels in sensory neurons that are pivotal in control of visceral sensation.

描述（由申请人提供）：A-激酶锚定蛋白（AKAP）通过将其分子成分聚集到离散的亚细胞微区域，来组织许多细胞内信号通路。一种这样的AKAP AKAP79/150与蛋白激酶A，蛋白激酶C（PKC），钙调蛋白（CAM），钙调蛋白（CAN），M型（KCNQ，KV7）K+通道，L-type Ca2+通道和G蛋白蛋白核的受体相互作用。已活化的T细胞（NFAT）转录因子家族的核因子已证明是活性依赖性调节基因转录的基因，包括中枢神经元和周围神经元在内。鼻子神经节的感觉神经元负责将传入从内脏器官发送到控制体器官的脑干。在该项目中，我们将通过NFATC1-C4成员研究KCNQ2-3通道的转录调控，这些神经元在上颈神经节（SCG）交感神经元中被NFATC1-C4成员，这些成员被细胞内Ca2+（Ca2+ I）信号激活，例如由活性依赖于活性依赖于活性的，这些信号。 We hypothesize the Ca2+i signals are sensed by CaN, anchored by AKAP79/150 to L-type CaV1.3 Ca2+ channels, followed by dephosphorylation of NFATs, their import into the nucleus, and up-regulation of M-channel gene expression that acts as a negative feedback on excitability in physiological and pathophysiological states.我们将使用各种尖端方法检验这一假设。我们进一步假设AKAP79/150通过各种对疼痛和炎症至关重要的GQ/11连锁受体类型来编排M电流的调节，这些受体对疼痛和炎症至关重要，包括Bradykinin，Par-2，组胺，血管紧张素II和嘌呤能受体。我们将确定哪种GQ/11耦合受体调节NG神经元，以及AKAP79/150在将受体耦合到M型通道中的作用。要使用的技术包括天然SCG和NG神经元的斑块夹电生理学，单个活细胞成像，共焦显微镜，荧光素酶基因转化蛋白分析，遗传性改变的小鼠以及活体动物的空气道。我们渴望在组织M型K+通道的转录调控方面发现赋予AKAP79/150的机制，并在对内脏感觉控制中关键的感官神经元中的这些通道划定受体信号。