Defining the circuit, synaptic, and molecular mechanisms linking intracellular Ca2+ release to learning using subcellularly-targeted manipulations and imaging techniques in dendrites in vivo

使用体内树突的亚细胞靶向操作和成像技术定义连接细胞内 Ca2 释放与学习的电路、突触和分子机制

• 批准号: 10502363
• 负责人: Justin O'Hare
• 金额: $ 13.62万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10502363
• 关键词: Action Potentials; Address; Alzheimer' s Disease; Animals; Apical; Area; Automobile Driving; BRAIN initiative; Behavior; Behavioral; Biophysics; Brain; Calcium; Cells; Color; Communities; Consultations; Dendrites; Development Plans; Electroporation; Endoplasmic Reticulum; Environment; Equipment; Fire - disasters; Foundations; Glutamates; Goals; Head; Hippocampus (Brain); Image; Imaging Techniques; In Vitro; Institutes; Intervention; Investigation; Knowledge; Laboratories; Learning; Light; Link; Literature; Logic; Memory; Mentorship; Mission; Molecular; Molecular Genetics; Monitor; Morphology; Mus; Neurodegenerative Disorders; Neuronal Plasticity; Neurons; Neurosciences; Output; Parkinson Disease; Pathway interactions; Pattern; Pharmacology; Play; Prevalence; Reporter; Reporting; Research; Research Activity; Resource Development; Role; Shapes; Supervision; Synapses; Synaptic plasticity; Testing; Work; autism spectrum disorder; awake; base; career development; cellular imaging; experience; experimental study; gain of function; hippocampal pyramidal neuron; imaging approach; improved; in vivo; innovation; insight; nervous system disorder; neuronal cell body; novel; optogenetics; place fields; post-doctoral training; postsynaptic; presynaptic; receptive field; relating to nervous system; spatiotemporal; subcellular targeting; tool; two-photon; virtual environment; virtual reality; way finding

Project Summary/Abstract Candidate Goals and Mission Relevance: The applicant’s broad, long-term objective is to investigate how high- (circuit/behavioral) and low- (subcellular/molecular) level organizational principles of the brain cooperate to drive learning. The proposed research activities will build a foundation for this long-term goal and, in so doing, will promote BRAIN 2025 Report goals by integrating new technological and conceptual approaches to causally link intracellular Ca2+ release (ICR) from endoplasmic reticulum (ER) to neural activity dynamics and behavior. Project description: Dendritic Ca2+ is central to neural plasticity mechanisms allowing animals to adapt to the environment. ICR has long been thought to shape these mechanisms. The applicant recently carried out the first investigation of ICR in mammalian neurons in vivo to uncover how this subcellular phenomenon shapes experience-dependent feature selectivity across the dendritic arbor of pyramidal neurons (PNs) in mouse hippocampal area CA1. This work raises important questions regarding when, where, and how ICR is engaged to support learning. The applicant will address these questions in the following Aims: Aim 1. Characterize plasticity-associated ER Ca2+ dynamics in dendrites in vivo (K99): To achieve this Aim, the applicant will perform simultaneous dual-color, dual-plane in vivo 2-photon imaging of cytosolic and ER- resident Ca2+ in dendrites of single CA1 PNs during head-fixed spatial navigation of novel virtual environments. Aim 2. Define the synaptic logic tying intracellular Ca2+ release to in vivo synaptic plasticity (K99/R00): The applicant will first create a novel molecular tool to optogenetically induce ICR (Aim 2.1; K99). The applicant will then combine this precise interventional tool with single-cell imaging, inducible blockade of presynaptic release, and optogenetic dampening of ICR to dissect the synaptic logic by which ICR participates in plasticity induction in behaving mice. (Aim 2.2; R00). Aim 3. Dissect excitatory circuit-molecular mechanisms driving intracellular Ca2+ release in vivo (R00): The candidate will optogenetically activate specific excitatory projections onto distinct dendritic compartments of single CA1PNs while monitoring ER Ca2+ dynamics in behaving mice. Local pharmacological manipulations will dissect contributions of the two canonical pathways that convert presynaptic excitatory input to postsynaptic ICR. Career development plan: The applicant will extend a highly complementary Co-Mentorship arrangement between Drs. Franck Polleux and Attila Losonczy who possess deep expertise in cellular/molecular/genetic and in vivo/behavioral approaches, respectively. The applicant will receive robust consultative support from Dr. Stefano Fusi of Columbia’s Center for Theoretical Neuroscience and Dr. Darcy Peterka, Director of Cellular Imaging at Columbia’s Zuckerman Institute. The applicant’s research and transition to independence will benefit from this strong mentorship team, state-of-the-art facilities, all necessary equipment, and numerous Professional Development resources offered through the Columbia Office of Postdoctoral Affairs, the Zuckerman Institute, and the BRAIN Initiative.

项目摘要/摘要 候选目标和使命相关性：申请人的广泛长期目标是调查高度 （电路/行为）和低 - （亚细胞/分子）大脑合作的组织原理以驱动 学习。拟议的研究活动将为这个长期目标奠定基础，以此为此 通过整合新的技术和概念方法来促进大脑2025报告目标 细胞内Ca2+释放（ICR）从内质网（ER）到神经活动动力学和行为。 项目描述：树突状CA2+对于允许动物适应的神经可塑性机制是核心 环境。长期以来，ICR一直被认为可以塑造这些机制。适当的最近进行了第一个 研究体内哺乳动物神经元中ICR的研究，以发现这种亚细胞现象的形状如何 小鼠锥体神经元（PNS）的树突状乔木的经验依赖性特征选择性 海马区CA1。这项工作提出了有关ICR何时，何时和如何参与的重要问题 支持学习。申请人将在以下目的中解决这些问题： 目的1。在体内树突中与塑性相关的ER Ca2+动力学表征（K99）：为此 AIM，申请人将在胞质和ER的体内同时执行双色双平面2光子成像 在新型虚拟环境的头部固定空间导航期间，单个CA1 PN的树突中的居民Ca2+。 AIM 2。定义合成逻辑绑定细胞内Ca2+释放到体内合成可塑性（K99/R00）： 申请人将首先创建一种新型的分子工具，以诱导ICR（AIM 2.1; K99）。申请人 然后将这种精确的介入工具与单细胞成像，可诱导的突触前封锁 释放ICR的释放和光遗传性减弱，以剖析ICR参与者可塑性的突触逻辑 行为小鼠的诱导。 （AIM 2.2; R00）。 目标3。解剖兴奋性电路分子机制，驱动细胞内Ca2+体内释放（R00）： 候选人将光遗传学将特定的令人兴奋的项目激活到不同的树突室 单个CA1PN在监视行为小鼠中的ER Ca2+动力学时。当地的药物操纵将 剖析两种规范途径的贡献，这些途径将突触前兴奋性输入转化为突触后ICR。 职业发展计划：申请人将扩展一项高度完善的会议安排 在DR之间。 Franck Polleux和Attila Losonczy在细胞/分子/遗传方面具有深厚的专业知识 体内/行为方法分别采用。申请人将获得Dr.博士的强大咨询支持。 哥伦比亚理论神经科学中心的Stefano Fusi和Cellular主任Darcy Peterka博士 在哥伦比亚的Zuckerman研究所成像。申请人的研究和过渡将受益 从这个强大的精通团队，最先进的设施，所有必要的设备和众多专业人士 通过哥伦比亚博士后事务办公室，扎克尔曼研究所提供的发展资源， 和大脑倡议。