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

• 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
• 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' 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

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.

摘要