Limbic Circuit Dysfunction in Offspring following Maternal Immune Activation

母体免疫激活后后代的边缘回路功能障碍

• 批准号: 9314190
• 负责人: John R Huguenard
• 金额: $ 23.73万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-15 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9314190
• 关键词: Acute; Address; Adolescence; Adolescent; Adult; Adult Children; Age-Months; Amygdaloid structure; Antiviral Agents; Axon; Behavior Disorders; Behavioral; Brain; Brain region; Cells; Child; Childhood; Cognitive; Comorbidity; Data; Defect; Depressed mood; Disease; Disease model; Distal; Electrophysiology (science); Employee Strikes; Environmental Risk Factor; Epilepsy; Etiology; Evaluation; Excitatory Synapse; Exposure to; Functional disorder; General Population; Generations; Glutamates; Hippocampus (Brain); Immune; Immune system; Immunohistochemistry; Individual; Infection; Inflammatory Response; Injection of therapeutic agent; Interneurons; Intervention; Investigation; Kinetics; Laboratories; Lasers; Lead; Life; Light; Literature; Location; Maps; Measures; Medial; Methods; Microcephaly; Modeling; Morphology; Mus; Neurodevelopmental Disorder; Neurons; Optics; Pathogenesis; Pattern; Phenotype; Physiologic pulse; Play; Poly I-C; Positioning Attribute; Predisposition; Prefrontal Cortex; Pregnancy; Probability; Public Health; Recruitment Activity; Reporting; Research; Research Project Grants; Rest; Risk; Risk Factors; Rodent; Role; Saline; Scanning; Seizures; Severities; Site; Slice; Social Interaction; Structural defect; Structure; Surveys; Synapses; Techniques; Temporal Lobe Epilepsy; Tissues; Viral; Work; Zika Virus; autism spectrum disorder; base; biocytin; calmodulin-dependent protein kinase II; cell type; emotional behavior; epidemiology study; excitatory neuron; experimental study; fetal; genetic risk factor; hippocampal pyramidal neuron; immune activation; improved; in vitro activity; mimetics; neural circuit; offspring; optogenetics; postsynaptic; pregnant; prenatal exposure; presynaptic; presynaptic neurons; response; risk sharing; sound; stereotypy; synaptic inhibition; targeted treatment; voltage clamp

PROJECT SUMMARY/ABSTRACT In the general population, less than 1% of children develop seizures, whereas over 30% of children with autism spectrum disorder (ASD) do so by adolescence. Maternal infection during pregnancy is a risk factor for both ASD and epilepsy, however we have limited understanding of how early-life immune insults alter neural circuitry implicated in both disorders. The medial prefrontal cortex (mPFC) is a brain region important for regulating diverse cognitive and emotional behaviors altered in ASD. We have obtained pilot data supporting altered mPFC circuit function after gestational exposure to a viral mimetic polyinosinic:polycytidylic acid (poly(I:C)). Using a high-throughput multisite field potential recording approach in acute mPFC brain slices, our preliminary results indicate that maternal immune activation (MIA) leads to persistent alterations in mPFC activity, at both presynaptic and postsynaptic loci, and an increased susceptibility to generation of epileptic activities in vitro. The proposed experiments will elucidate the precise cell type- and lamina-specific alterations underlying changes in network excitability. Aim 1 of this proposal will identify changes to functional intracortical connectivity within the mPFC. Pregnant dams will be treated with saline or poly(I:C) to mount an acute antiviral inflammatory response. Acute brain slices containing mPFC will be prepared from adult offspring at three to four months of age. Laser scanning photostimulation based on patterned glutamate uncaging will be used to map excitatory synaptic connectivity within the mPFC between photo-excited presynaptic neurons and individual postsynaptic pyramidal and interneurons. Changes to laminar-specific inputs to excitatory and inhibitory cells types following MIA will be assessed by intracellular voltage-clamp recordings of synaptic activity. Immunohistochemical methods will confirm cell type, location and morphology and will be used to identify structural defects in lamination. Aim 2 will evaluate alterations in the recruitment of distal inputs to the mPFC from two regions highly implicated in the initiation and spread of limbic seizures, the basolateral amygdala (BLA) and ventral hippocampus (vHC). AAV-CamKII-ChR2/eYFP will be used to target expression of channelrhodopsin-eYFP or eYFP alone to excitatory neurons in these extra-prefrontal structures. Postsynaptic currents in mPFC in L2/3 and L5 neurons will be measured in response to optogenetic activation of BLA and vHC terminals, respectively, and changes in excitatory and inhibitory components assessed. Together these aims address circuit intrinsic and extrinsic changes that may contribute to the shared pathogenesis of ASD and epilepsy. Information gained from the interrogation of mPFC circuitry in an established MIA model will identify neuropathological alterations that might be targeted for therapies.

项目概要/摘要 在一般人群中，不到 1% 的儿童会出现癫痫发作，而超过 30% 的儿童患有自闭症 青春期母亲感染谱系障碍 (ASD) 是这两种疾病的危险因素。 自闭症谱系障碍（ASD）和癫痫，然而，我们对生命早期免疫损伤如何改变神经系统的了解有限。 内侧前额叶皮层 (mPFC) 是与这两种疾病有关的重要大脑区域。 调节自闭症谱系障碍中不同的认知和情绪行为我们已经获得了支持的试点数据。 妊娠期暴露于病毒模拟物聚肌苷：聚胞苷酸后 mPFC 回路功能发生改变 （poly(I:C)）在急性 mPFC 脑切片中使用高通量多位点场电位记录方法，我们的研究 初步结果表明母体免疫激活（MIA）导致 mPFC 持续改变 突触前和突触后位点的活动，以及对癫痫发作的易感性增加 拟议的实验将阐明精确的细胞类型和层特异性改变。 该提案的目标 1 将确定皮质内功能的变化。 mPFC 内的连接将用盐水或聚 (I:C) 进行处理，以安装急性抗病毒药物。 将从成年后代中制备含有 mPFC 的急性炎症反应。 四个月大时，将使用基于图案化谷氨酸解笼锁的激光扫描光刺激。 绘制 mPFC 内光激发突触前神经元和 个体突触后锥体和中间神经元的层状特异性输入的变化。 MIA 后的抑制性细胞类型将通过突触的细胞内电压钳记录进行评估 免疫组织化学方法将确认细胞类型、位置和形态，并将用于 识别层压中的结构缺陷，目标 2 将评估远端输入募集的变化。 来自两个区域的 mPFC 与边缘癫痫发作的起始和扩散高度相关，即基底外侧区域 杏仁核 (BLA) 和腹侧海马 (vHC) 将用于靶向表达。 视紫红质通道蛋白-eYFP 或单独的 eYFP 作用于这些前额叶外结构中的兴奋性神经元。 将测量 L2/3 和 L5 神经元 mPFC 中的突触后电流以响应光遗传学激活 分别评估 BLA 和 vHC 末端的变化，并评估兴奋性和抑制性成分的变化。 这些目标共同解决了电路内在和外在的变化，这些变化可能有助于共享 自闭症谱系障碍 (ASD) 和癫痫的发病机制。 建立的 MIA 模型将识别可能作为治疗目标的神经病理学改变。