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Probing ASIC1 function in vivo using novel genetic tools

使用新型遗传工具探测 ASIC1 体内功能

基本信息

批准号：
9109049
负责人：
CECILIA M CANESSA
金额：
$ 24.98万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-10 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9109049
关键词：
ASIC channel Acids Action Potentials Address Affect Amygdaloid structure Anxiety Area Astrocytes Behavior Behavior Therapy Behavioral Binding Brain Cell Nucleus Cells Conditioned Reflex Defect Development Drops Experimental Models Extracellular Space Family Fright Genetic Glutamates Goals Health Individual Injection of therapeutic agent Interneurons Invertebrates Ion Channel Ischemia Knock-out Knockout Mice Learning Light Lighting Location Memory Methods Modeling Mus Neurons Output Parvalbumins Perfusion Phase Physiological Population Protocols documentation Proton Pump Protons Reporting Resolution Rest Role Sodium Channel Somatostatin Staging Stimulus Stream Structure Substance abuse problem Synaptic plasticity Vertebrates Viral Vector Work base behavioral response cell type conditioned fear desensitization epithelial Na+ channel genetic approach genetic inhibitor in vivo inhibitor/antagonist interstitial learned behavior nanobodies nervous system disorder neuronal circuitry novel optogenetics recombinase research study response signal processing spatiotemporal stem tool

项目摘要

DESCRIPTION (provided by applicant): ASIC or Acid Sensing Ion Channel is a proton-gated sodium channel that belongs to the large family of Deg/ENaC ion channels expressed in vertebrates and invertebrates. ASIC has been implicated in various neurological disorders though their physiological roles remain poorly defined. Slow progress elucidating ASIC functions stems in part from technical difficulties of changing the interstitial pH in selected areas of the brain. To date most attempts to activate ASIC have used perfusion of acid solutions or induction of ischemia; however such maneuvers have low spatial and temporal control of the concentration of protons and also tend to suppress ASIC activity because they induce desensitization. To overcome this problem we have implemented an optogenetic-based method that acidifies selected areas of the extracellular space in the CNS with high temporospatial resolution enabling activation of ASIC in any structure of the brain. The method consists on expressing the light-driven proton pump ArchT in astrocytes that upon illumination extrude protons lowering the pH surrounding nearby neurons. The acidification activates ASIC and initiates spiking of action potentials. Another hurdle in the field is the ubiquitous expression of ASIC in the CNS - almost all neurons independent of location or functional specialization express ASIC; thus, all the neurons in a microcircuit are affected by drops in pH, making difficult to tease out the contribution of individual neurons to a behavioral response. To address this problem we have developed a genetically encoded specific inhibitor of ASIC; it consists of a `nanobody' that binds to the aminoterminus and inhibits channel activity. This proposal aims to validate the newly developed tools by examining the contribution of ASIC in a well-defined neuronal microcircuit; specifically, we will investigate how ASIC modulates processing of signals encoding fear learning and memory in the basolateral amygdala. Although ASIC is present in most areas of the brain, we will focus in the amygdala because previous reports suggest that it modulates fear responses, albeit the cell types involved and the mechanism remain largely unknown. The work will prove the value of these novel genetic approaches to studying ASIC in behaviors and sets the stage for a next phase of exploration of the physiological and pathological roles of these channels in the mammalian brain.