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.

项目摘要 皮质亚区通常与多种行为功能有关，但其机制尚不清楚。 这些区域编码如此多样的信息。前扣带皮层（ACC）是情绪调节所必需的。 加工和社会认知，但它如何编码刺激相关的两个过程是未知的。的目标 这个建议是为了了解ACC电路的功能异质性，以及这如何影响编码。 不同的刺激和认知功能。在目的1中，我们将确定ACC抑制剂的功能异质性。 在社交和类似焦虑的行为中的回路。为了监测中间神经元的活动，我们将首先注射一个腺- 相关病毒（AAV），其以Cre依赖性方式表达GCaMP 6 f（一种荧光Ca 2+指示剂）， 生长抑素（SST）或小白蛋白（PV）-cre转基因小鼠ACC。接下来，我们将使用3D打印的 微型微型镜进行体内单细胞分辨率钙成像在生长抑素或 ACC中的小清蛋白中间神经元，以研究它们的功能异质性，同时小鼠执行任务， 分析焦虑样行为、一般社交能力和社交新奇感。在目标2中，我们将确定是否具体 ACC中的兴奋性锥体细胞（Pyr）群体编码特定的行为刺激。第一，瞄准 为了在ACC中表达Pyr的特定群体，我们将注射表达Cre重组酶的AAV， 逆行标记细胞进入对侧ACC、丘脑前核或压后皮质。 接下来，我们将注射以cre依赖性方式表达GCaMP 6的AAV以标记ACC中的Pyr， 投射到对侧ACC、丘脑前核或压后皮质。三周后，我们 将使用目标1中描述的方法来监测特定Pyr种群在特定时期的活动， 行为。我们还将确定ACC中VIP中间神经元的突触输入的层状来源。 为此，我们将首先将VIP-cre小鼠与cre依赖性荧光报道细胞系杂交。接下来，我们将注射一个AAV 表达ChETA，一种光激活通道，进入对侧ACC，丘脑前核或 压后皮质病毒注射后四周，我们将联合收割机结合急性期的电生理记录， 具有光遗传学刺激的脑切片，以表征VIP特异性投射的层状组织 除了确定这些输入的连接概率外，我们还将关联 皮质深度的索马的形态，和电生理特性和分子标记。 虽然皮层网络的简化模型已经建立了一个框架来理解它们的功能， 功能，现在很明显，要了解皮层网络在复杂的适应性行为中的作用， 优化的模型将需要结合这些电路的功能异质性。完成这些 目的是剖析ACC中抑制回路的功能异质性。