Functional heterogeneity of vasoactive intestinal peptide-expressing interneurons in the anterior cingulate cortex

前扣带皮层表达血管活性肠肽的中间神经元的功能异质性

• 批准号: 10612970
• 负责人: Alberto Cruz-Martin
• 金额: $ 41.25万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612970
• 关键词: 3D Print; Acute; Adaptive Behaviors; Address; Anterior; Anterior Nuclear Group; Anti-Anxiety Agents; Anxiety; Area; Automobile Driving; Behavior; Behavior monitoring; Behavioral; Behavioral trial; Biological Assay; Brain; Calcium; Categories; Cells; Cerebral cortex; Classification; Complex; Contralateral; Data; Decision Making; Dependovirus; Electrophysiology (science); Emotional; Enterobacteria phage P1 Cre recombinase; Exhibits; Goals; Heterogeneity; Image; Implant; Individual; Injections; Interneurons; Label; Light; Maps; Methods; Microscope; Modeling; Molecular; Monitor; Morphology; Mus; Neurons; Parvalbumins; Pattern; Population; Positioning Attribute; Probability; Process; Property; Pyramidal Cells; Rabies; Reporter; Resolution; Rodent; Role; Slice; Social Behavior; Somatostatin; Source; Specificity; Stimulus; Synapses; Testing; Thalamic Nuclei; Transgenic Mice; Vasoactive Intestinal Peptide; Viral; Work; anxiety-like behavior; cingulate cortex; classification algorithm; cognitive function; emotion regulation; emotional behavior; excitatory neuron; implantation; in vivo; individual response; insight; miniaturize; molecular marker; neural; neuronal cell body; optogenetics; presynaptic; response; social; social cognition

PROJECT SUMMARY Cortical subregions are often implicated in a variety of behavioral functions, but it is not well understood how these areas encode such diverse information. The anterior cingulate cortex (ACC) is necessary for emotional processing and social cognition, but how it encodes stimuli relevant to both processes is unknown. The goal of this proposal is to understand the functional heterogeneity of ACC circuits, and how this impacts encoding of diverse stimuli and cognitive function. In Aim 1, we will determine the functional heterogeneity of ACC inhibitory circuits during social and anxiety-like behaviors. To monitor interneuron activity, we will first inject an adeno- associated virus (AAV) that expresses GCaMP6f, a fluorescent Ca2+ indicator, in a Cre-dependent manner into the ACC of somatostatin (SST)- or parvalbumin (PV)-cre transgenic mice. Next, we will use the 3D-printed miniature miniscopes to perform in vivo single-cell resolution calcium imaging in either somatostatin or parvalbumin interneurons in the ACC to investigate their functional heterogeneity while mice performed tasks to assay anxiety-like behaviors, general sociability and social novelty. In Aim 2, we will determine whether specific populations of excitatory pyramidal cells (Pyr) in the ACC encode particular behavioral stimuli. First, to target specific populations of Pyr in the ACC, we will inject an AAV that expresses Cre recombinase and can retrogradely label cells into either the contralateral ACC, anterior thalamic nucleus, or the retrosplenial cortex. Next, we will inject an AAV that expresses GCaMP6 in a cre-dependent manner to label Pyr in the ACC and project to either the contralateral ACC, anterior thalamic nucleus or retrosplenial cortex. Three weeks later, we will use approaches described in Aim 1 to monitor the activity of specific populations of Pyr during particular behaviors. We will also determine the laminar sources of synaptic input to VIP interneurons in the ACC. To test this, we will first cross VIP-cre mice to a cre-dependent fluorescent reporter line. Next, we will inject an AAV expressing ChETA, a light-activated channel, into the contralateral ACC, anterior thalamic nucleus or retrosplenial cortex. Four weeks after viral injections, we will combine electrophysiological recordings in acute brain slices with optogenetic stimulation to characterize the laminar organization of specific projections unto VIP interneurons in the ACC. In addition to determining the connectivity probability of these inputs, we will correlate the cortical depth of the soma to morphological, and electrophysiological properties and molecular markers. While reductionist and simplified models of cortical networks have established a framework to understand their function, it is now evident that to understand the role of cortical networks in complex, adaptive behavior, optimized models will need to incorporate the functional heterogeneity of these circuits. Completion of these aims will dissect the functional heterogeneity of inhibitory circuits in the ACC.

项目总结 大脑皮层亚区通常与多种行为功能有关，但目前还不清楚是如何实现的 这些区域编码了如此多样化的信息。前扣带回皮质(ACC)是情绪调节所必需的 加工和社会认知，但它如何编码与这两个过程相关的刺激尚不清楚。的目标是 这项建议是为了了解ACC电路的功能异构性，以及这如何影响编码 不同的刺激和认知功能。在目标1中，我们将确定ACC抑制物的功能异质性 在社交和类似焦虑的行为中循环。为了监测神经元间的活动，我们首先要注射一个腺体- 相关病毒(AAV)，以Cre依赖的方式表达荧光钙指示剂GCaMP6f，进入 生长抑素(SST)或小白蛋白(PV)-cre转基因小鼠的ACC。接下来，我们将使用3D打印的 在生长抑素或生长抑素中进行体内单细胞分辨率钙成像的微型微型显微镜 在小鼠执行任务时，研究ACC中的小白蛋白中间神经元的功能异质性 测试焦虑样行为、一般社交能力和社交新颖性。在目标2中，我们将确定是否具体 ACC中的兴奋性锥体细胞群(PYR)编码特定的行为刺激。第一，对准目标 在ACC中，我们将注射一种表达Cre重组酶并能 逆行标记细胞进入对侧ACC、丘脑前核或脾后皮质。 接下来，我们将注射一种以依赖于cre的方式表达GCaMP6的AAV来标记ACC中的PYR和 投射到对侧ACC、丘脑前核或脾后皮质。三周后，我们 将使用目标1中描述的方法来监测特定群体在特定时期的活动 行为。我们还将确定ACc内VIP中间神经元的突触输入的层流来源。为了测试 为此，我们将首先将VIP-cre小鼠与cre依赖的荧光报告系杂交。接下来，我们将注射一种AAV Cheta，一种光激活通道，表达到对侧ACC、丘脑前核或 脾后皮质。病毒注射四周后，我们将结合急性脑出血患者的电生理记录 光遗传刺激脑片表征VIP特定投射的层状组织 ACC中的中间神经元。除了确定这些输入的连接概率外，我们还将关联 胞体皮质深度对形态、电生理特性和分子标记的影响。 而大脑皮层网络的简化论者和简化模型已经建立了一个框架来理解他们的 功能，现在很明显，要了解皮质网络在复杂的适应性行为中的作用， 优化的模型需要包含这些电路的功能异构性。完成这些工作 AIMS将剖析ACC中抑制回路的功能异质性。