Neurobiology of loss: role of microglia

损失的神经生物学：小胶质细胞的作用

• 批准号: 10593176
• 负责人: Marissa Ann Smail
• 金额: $ 2.3万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-09-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10593176
• 关键词: Amygdaloid structure; Automobile Driving; Behavior; Behavioral; Bioinformatics; Cannulas; Cells; Cognitive; Confocal Microscopy; Data; Depressed mood; Development; Down-Regulation; Drug Delivery Systems; Emotions; Environment; Event; Excision; Exhibits; Exposure to; Fluorescence-Activated Cell Sorting; Genes; Genetic Transcription; Goals; Homeostasis; Hormones; Housing; Human; Hyperphagia; Impairment; Implant; Income; Intervention; Laboratories; Link; MAP Kinase Gene; MAP3K8 gene; Maintenance; Major Depressive Disorder; Mental Depression; Microglia; Molecular; Molecular Profiling; Monitor; Morphology; Nature; Negative Valence; Neuroanatomy; Neurobiology; Neurons; Pathology; Pathway interactions; Persons; Phenotype; Phosphotransferases; Physiological; Physiology; Play; Poverty; Process; Proteins; Proteomics; Protocols documentation; RNA; Rattus; Reaction; Research; Research Domain Criteria; Rewards; Rodent; Role; Signal Pathway; Signal Transduction; Source; Stimulus; Stress; Symptoms; Techniques; Testing; Time; Weight Gain; Withdrawal; activity marker; antagonist; behavior test; behavioral outcome; behavioral pharmacology; biological adaptation to stress; cell type; coping; cytokine; environmental enrichment for laboratory animals; experience; experimental study; innovation; insight; maladaptive behavior; male; molecular marker; morphometry; multiple omics; neurotransmission; osmotic minipump; pharmacologic; physical conditioning; psychologic; receptor; response; social; social relationships; therapeutic target; transcriptome sequencing

Project Summary Psychological loss occurs when one is deprived of something valuable, such as social relationships, physical health, or financial stability. These experiences often lead to depression-like symptoms such as amotivation, social withdrawal, weight gain and hypoactive stress responses. While loss experiences are common, little is known about the molecular mechanisms driving loss symptomology. Our laboratory uses enrichment removal (ER) to simulate the experience of loss in rats. In this protocol, rats are first housed in positive, highly stimulating environmental enrichment (EE) for a month, and then moved to single housing with no further stimuli. This removal alters how the rats respond to their environment, producing behaviors and physiology that specifically resemble loss in humans, including increased passive coping, weight gain, and hypoactive stress responses. We used an innovative discovery-based multi-omics approach to investigate ER molecular mechanisms in the basolateral amygdala (BLA), a traditionally stress promoting region that is differentially activated in EE and ER. Using a combination of RNA sequencing, proteomics, and kinomics, we identified a unique microglia phenotype in ER. We hypothesize that ER decreases signaling in BLA microglia, impairing their ability to regulate nearby neurons and dampen stress-promoting BLA signals. This dysregulated microglia signaling is supported on all three omics platforms and is expected to contribute to ER behavioral and physiological phenotypes. Additional analyses revealed that MAP3K8, a microglia-specific kinase, is a signaling hub in this phenotype and could be a potential target to ameliorate ER phenotypes. The objective of this proposal is to further define BLA microglia phenotypes in ER and investigate the role of the signaling hub MAP3K8 in ER behavioral and physiological phenotypes. Specific Aim 1 will use fluorescence activated cell sorting (FACS) to isolate BLA microglia, followed by qPCR to determine microglia-specific expression of key genes identified in our preliminary analyses. Aim 1 will also use confocal microscopy and quantitative morphometry to determine ER-related microglia morphology and expression of MAPK activity markers. These well-established, cell type specific studies are necessary to clearly define the specific role of microglia, as opposed to other cell types, in ER phenotypes. Specific Aim 2 will use behavioral pharmacology to examine the role of MAP3K8 in ER behavioral and physiological phenotypes. We will implant BLA-targeted cannula attached to osmotic minipumps containing a MAP3K8 antagonist prior to the ER paradigm and behavioral testing. Differential timing of drug delivery will allow us to investigate both the necessity and sufficiency of MAP3K8 signaling. Overall, the findings of these studies will define the role of BLA microglia and MAP3K8 signaling in ER phenotypes, and provide possible insight into the negative consequences of loss in humans.

项目摘要 当一个人被剥夺了一些有价值的东西时，比如社会关系、身体上的， 健康或财务稳定。这些经历往往会导致抑郁样症状，如动力不足， 社交退缩、体重增加和应激反应减退。虽然损失的经验是常见的， 关于分子机制的知识驱动损失病理学。我们的实验室使用浓缩去除 (ER)来模拟老鼠失去亲人的经历。在该方案中，首先将大鼠圈养在阳性、高度 刺激环境丰富（EE）一个月，然后搬到单一的住房，没有进一步的 刺激。这种移除改变了大鼠对环境的反应，产生了行为和生理学， 特别是类似于人类的损失，包括增加被动应对，体重增加和低活动压力 应答我们使用了一种创新的基于发现的多组学方法来研究ER分子 基底外侧杏仁核（BLA）是一个传统的压力促进区域， 在EE和ER中激活。使用RNA测序，蛋白质组学和运动组学的组合，我们确定了一个 ER中独特的小胶质细胞表型。我们假设ER减少了BLA小胶质细胞的信号传导， 它们调节附近神经元和抑制促进压力的BLA信号的能力。这种失调的小胶质细胞 所有三个组学平台都支持信号传导，预计将有助于ER行为和 生理表型进一步的分析显示，MAP3K8是一种小胶质细胞特异性激酶， 在这种表型的信号枢纽，并可能是一个潜在的目标，以改善ER表型。的目标 本研究旨在进一步确定ER中BLA小胶质细胞的表型，并研究信号中枢的作用 ER行为和生理表型中的MAP3K8。Specific Aim 1将使用荧光激活细胞 分选（FACS）以分离BLA小胶质细胞，然后进行qPCR以确定关键的免疫球蛋白的小胶质细胞特异性表达。 在我们的初步分析中发现的基因。Aim 1还将使用共聚焦显微镜和定量 形态计量法测定ER相关小胶质细胞形态和MAPK活性标志物的表达。这些 有必要进行成熟的细胞类型特异性研究，以明确小胶质细胞的特定作用， 与其他细胞类型相反，在ER表型中。具体目标2将使用行为药理学来检查 MAP3K8在ER行为和生理表型中的作用。我们将植入BLA靶向套管 在ER模式和行为模式之前，将其连接到含有MAP3K8拮抗剂的渗透微泵上， 试验.不同的给药时间将使我们能够研究的必要性和充分性， MAP3K8信号传导。总的来说，这些研究的结果将确定BLA小胶质细胞和MAP3K8的作用。 ER表型中的信号传导，并提供对人类损失的负面后果的可能见解。

项目成果

Cellular, molecular, and therapeutic characterization of pilocarpine-induced temporal lobe epilepsy.

• DOI: 10.1038/s41598-021-98534-3
• 发表时间: 2021-09-27
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Henkel ND;Smail MA;Wu X;Enright HA;Fischer NO;Eby HM;McCullumsmith RE;Shukla R
• 通讯作者: Shukla R

Differential vulnerability of anterior cingulate cortex cell types to diseases and drugs.

前扣带回皮质细胞类型对疾病和药物的差异脆弱性。

• DOI: 10.1038/s41380-022-01657-w
• 发表时间: 2022-10
• 期刊: MOLECULAR PSYCHIATRY
• 影响因子: 11
• 作者: Smail, Marissa A.;Chandrasena, Sapuni S.;Zhang, Xiaolu;Reddy, Vineet;Kelley, Craig;Herman, James P.;Sherif, Mohamed;McCullumsmith, Robert E.;Shukla, Rammohan
• 通讯作者: Shukla, Rammohan