Fluorescent Indicators for Imaging Synaptic Zinc in Cortical Sound Processing

用于皮质声音处理中突触锌成像的荧光指示剂

基本信息

批准号：
10709627
负责人：
Huiwang Ai
金额：
$ 66.88万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10709627
关键词：
Address Affinity Alzheimer&apos s Disease Animals Auditory area Binding Brain Cells Cellular Membrane Chelating Agents Color Cytosol Development Directed Molecular Evolution Engineering Epilepsy Frequencies Generations Genes Hearing problem Image Imaging Device Imaging technology In Vitro Ischemia Knockout Mice Link Location Membrane Mental Depression Mus Neurobiology Neuromodulator Neurons Neurotransmitters Oranges Organelles Outcome Productivity Property Proteins Research Personnel Role SLC30A3 gene Sensory Shapes Signal Transduction Specificity Stimulus Stroke Synapses Synaptic Vesicles Testing Time Transgenic Mice Variant Vesicle Viral Vector Zinc auditory processing awake cell type extracellular fluorescence imaging imaging capabilities improved in vivo information processing innovation knockout gene nervous system disorder neurotransmission new technology novel presynaptic protein transport public health relevance response sensory cortex sensory stimulus sound sound frequency spatiotemporal synergism trafficking two-photon

项目摘要

Abstract Although the importance of synaptic Zn2+, as an emerging neuromodulator throughout the brain, has been widely appreciated, the dynamics of synaptic Zn2+ release in response to naturally occurring stimuli remains largely elusive. Genetically encoded Zn2+ indicators (GEZIs) derived from fluorescent proteins are popular tools for imaging Zn2+ in the cytosol and intracellular organelles. However, fluorescence imaging of Zn2+ secretion in the brain in live animals has not yet been achieved due to the limitations of current GEZIs (e.g., insufficient extracellular membrane localization, mismatching affinity, and/or inadequate dynamic range and photostability). This interdisciplinary multi-PI 4-year R01 project, led by Dr. Huiwang Ai with expertise in genetically encoded indicators and fluorescence imaging and Dr. Thanos Tzounopoulos with expertise in studying the role of Zn2+ in auditory processing, aims to (1) develop a new generation of GEZIs to address the hurdles for imaging secreted Zn2+ in the brain in vivo, and (2) integrate the new GEZIs with our innovative ZnT3 cKO mice, which, for the first time, allow for Cre-dependent expression of exogenous genes in ZnT3-expressing neurons and Dre-dependent region- and cell type-specific conditional ZnT3 gene knockout, to identify the cell- and circuit-specificity of Zn2+ dynamics that shape cortical sound processing. The project will lead to a novel capability of imaging synaptically released Zn2+ in the brain in awake behaving animals. Our innovative strategy to optimize the exoplasmic location of GEZIs may be generalized to enhance other genetically encoded indicators. Furthermore, because synaptic Zn2+ is a potent modulator throughout the cortex, our findings on Zn2+ dynamics in the primary auditory cortex (A1) during sound processing will improve the understanding of the roles of synaptic Zn2+ in cortical information processing beyond sensory cortices. We expect our studies to catalyze an extensive array of studies on Zn2+-related neurobiology and neurological diseases.

抽象的尽管突触Zn2+的重要性，但作为整个大脑的新兴神经调节剂值得赞赏的是，突触Zn2+释放的动力学响应天然刺激仍然很大程度上仍然存在难以捉摸。从荧光蛋白得出的遗传编码的Zn2+指标（GEZIS）是流行的工具在细胞质和细胞内细胞器中成像Zn2+。但是，Zn2+分泌的荧光成像由于当前GEZI的局限性，活动物的大脑尚未实现（例如，不足细胞外膜定位，不匹配亲和力和/或动态范围和光稳定性不足）。这个由Huiwang AI博士领导的跨学科的多PI 4年R01项目，具有遗传编码的专业知识指标和荧光成像和Thanos tzounopoulos博士在研究Zn2+在研究中的作用方面具有专业知识听觉处理，旨在（1）开发新一代的GEZI，以解决成像分泌的障碍体内大脑中的Zn2+，（2）将新的Gezis与我们的创新Znt3 CKO小鼠整合在一起，这是第一个时间，允许在表达Znt3的神经元和DRE依赖性中外源基因的CRE依赖性表达区域和细胞类型特异性条件Znt3基因敲除，以识别Zn2+的细胞和电路特异性塑造皮质声音处理的动力学。该项目将导致成像在大脑中突触释放的Zn2+的新颖能力。动物。我们优化GEZIS外质位置的创新策略可能会推广以增强其他遗传编码的指标。此外，由于突触Zn2+是整个过程中的有效调节器皮层，我们在声音处理过程中主听觉皮层（A1）中有关Zn2+动力学的发现将改善突触Zn2+在感觉皮层以外的皮质信息处理中的作用的理解。我们期望我们的研究能够催化有关Zn2+相关神经生物学和神经学的广泛研究疾病。