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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要皮质次区域通常与多种行为功能有关，但目前尚不清楚它是如何发挥作用的这些区域编码如此多样化的信息。前扣带皮层 (ACC) 对于情感表达是必需的处理和社会认知，但它如何编码与这两个过程相关的刺激尚不清楚。目标是该提案旨在了解 ACC 电路的功能异质性，以及这如何影响编码多样化的刺激和认知功能。在目标 1 中，我们将确定 ACC 抑制的功能异质性社交和焦虑类行为期间的回路。为了监测中间神经元活动，我们首先注射腺- 相关病毒 (AAV) 以 Cre 依赖性方式表达 GCaMP6f（一种荧光 Ca2+ 指示剂）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打印微型显微镜可在生长抑素或生长抑素中进行体内单细胞分辨率钙成像 ACC 中的小清蛋白中间神经元在小鼠执行任务时研究其功能异质性分析焦虑样行为、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。首先，要瞄准目标 ACC 中 Pyr 的特定群体，我们将注射表达 Cre 重组酶的 AAV，并且可以将细胞逆行标记到对侧 ACC、丘脑前核或压后皮质。接下来，我们将注入一个以依赖于 cre 的方式表达 GCaMP6 的 AAV，以在 ACC 中标记 Pyr 并投射到对侧 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时期 Pyr 特定群体的活动行为。我们还将确定 ACC 中 VIP 中间神经元突触输入的层流来源。测试为此，我们首先将 VIP-cre 小鼠与依赖于 cre 的荧光报告基因系杂交。接下来，我们将注入AAV 将 ChETA（一种光激活通道）表达到对侧 ACC、丘脑前核或压后皮质。病毒注射后四个星期，我们将结合急性期的电生理记录通过光遗传学刺激进行脑切片，以表征 VIP 特定投影的层状组织 ACC 中的中间神经元。除了确定这些输入的连接概率之外，我们还将关联体细胞的皮质深度与形态学、电生理学特性和分子标记有关。虽然皮层网络的还原论和简化模型已经建立了一个框架来理解它们功能，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，优化模型需要考虑这些电路的功能异质性。完成这些目标将剖析 ACC 中抑制回路的功能异质性。