Activity and connectivity of hippocampal newborn neurons underlie alcohol withdrawal-associated syndromes

海马新生神经元的活动和连接是酒精戒断相关综合征的基础

• 批准号: 10711653
• 负责人: Hoonkyo Suh
• 金额: $ 63.26万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711653
• 关键词: Alcohol Withdrawal Seizures; Alcohol abuse; Alcohol dependence; Alcohol withdrawal syndrome; Anatomy; Brain; Cells; Chronic; Color; Dangerousness; Data; Development; Epilepsy; Equilibrium; Foundations; Generalized seizures; Genes; Genetic Epistasis; Goals; Hippocampus; Hour; Hyperactivity; Image; Imaging Device; Longitudinal Studies; Magnetic Resonance Imaging; Maps; Mediating; Methods; Modality; Molecular; Neurons; Newborn Infant; Outcome; Parvalbumins; Pathologic; Play; Predisposition; Rabies virus; Recurrence; Relapse; Resolution; Role; Seizures; Series; Severities; Signal Transduction; Symptoms; Synapses; Syndrome; Testing; Time; Tonic - clonic seizures; Vertebral column; Visualization; Wheat Germ Agglutinins; alcohol effect; alcohol exposure; alcohol response; cell type; designer receptors exclusively activated by designer drugs; differential expression; excitatory neuron; experimental study; granule cell; hippocampal pyramidal neuron; imaging modality; inhibitory neuron; neural; neural circuit; neurogenesis; newborn neuron; novel; novel therapeutic intervention; postsynaptic neurons; presynaptic; single-cell RNA sequencing; transcriptome; transmission process

Alcohol withdrawal (AW) after chronic alcohol exposure (AE) produces a series of symptoms. Among them, generalized tonic-clonic seizures are the most severe and dangerous symptom. The severity and susceptibility to relapse, and perpetuation of alcohol abuse underscore the urgent need to understand mechanisms underlying alcohol dependence and AW in order to develop new therapeutic strategies to intervene and treat AW-associated syndromes such as seizures. In this application, we will test the novel hypothesis that activity and connectivity of hippocampal newborn dentate granule cells (DGCs) underlie AW-associated seizures. DGCs are principal excitatory neurons that are continuously produced and integrate into hippocampal neural circuits, and altered hippocampal neurogenesis has been implicated in seizures. Our previous studies have revealed the essential roles of hippocampal newborn DGCs in the expressions of AW-associated seizures. AE reduced spine formation while AW increased synaptic connectivity of hippocampal newborn DGCs. Our rabies virus-mediated retrograde tracing study discovered altered neuronal connectivity of hippocampal newborn DGCs with both excitatory and inhibitory neurons during AW seizures. Moreover, our functional study with a DREADD (Designer Receptors Exclusively Activated by Designer Drugs) method demonstrated that activity of hippocampal newborn neurons plays an essential role in the expression of AW-associated seizures. These observations provided the theoretical foundation for our hypothesis that altered neuronal connectivity and activity of hippocampal newborn DGCs disrupts the balance of excitatory and inhibitory (E/I) signals, ultimately leading to AW-associated seizures. Thus, the central goal of this proposal is to use novel mapping methods, imaging tools, and cellular and molecular approaches in order to understand activity and connectivity of hippocampal neural circuits that are responsible for AW-associated seizures. In Aim 1, we will determine whether AW alters neuronal and functional connectivity of DGCs by using a rabies virus- and wheat germ agglutinin (WGA)-mediated retrograde and anterograde tracing methods, respectively. We will also use multiple DREADDs and assess the essential role of de novo neural circuits formed between hippocampal newborn DGCs and input neurons in AW-associated seizures. Aim 2, using Ca2+ imaging and various magnetic resonance imaging (MRI) modalities that allows us for longitudinal studies, we will determine activity and connectivity of hippocampal and global neural circuits underlying AW-associated seizures. In Aim 3, we will identify and validate transcriptome that may underlie altered synaptic and neuronal connectivity of hippocampal newborn DGCs in response to AE and AW. The usage of single cell RNA sequencing will allow us to not only register differentially expressed genes to cell type-specific manner, but determine transcriptomes that distinguish pathological newborn DGCs from normal DGCs during AE and AW. Altogether, our proposal will dissect the molecular, cellular, and neural circuitry mechanisms by which hippocampal newborn DGCs underlie AW-associated seizures.

慢性酒精暴露（AE）后的酒精戒断（AW）会产生一系列症状。他们之中， 全身强直阵挛性癫痫发作是最严重和最危险的症状。严重性和敏感性 酗酒的复发和长期存在强调了迫切需要了解其背后的机制 酒精依赖和 AW，以开发新的治疗策略来干预和治疗 AW 相关的 癫痫等综合症。在此应用程序中，我们将测试新的假设，即活动和连接性 海马新生儿齿状颗粒细胞 (DGC) 是 AW 相关癫痫发作的基础。 DGC 是主要的 兴奋性神经元不断产生并整合到海马神经回路中，并改变 海马神经发生与癫痫发作有关。我们之前的研究已经揭示了重要的 海马新生儿 DGC 在 AW 相关癫痫发作表达中的作用。 AE 减少脊柱形成 而 AW 增加了海马新生 DGC 的突触连接。我们的狂犬病病毒介导的逆行 追踪研究发现海马新生儿 DGC 的神经元连接发生改变，兴奋性和 AW 癫痫发作期间的抑制性神经元。此外，我们使用 DREADD（设计受体）进行功能研究 由设计药物独家激活）方法证明海马新生神经元的活性 在 AW 相关癫痫发作的表达中起重要作用。这些观察结果提供了理论依据 为我们改变海马新生儿 DGC 的神经元连接和活性的假设奠定了基础 破坏兴奋性和抑制性 (E/I) 信号的平衡，最终导致与 AW 相关的癫痫发作。因此， 该提案的中心目标是使用新颖的绘图方法、成像工具以及细胞和分子 方法以了解负责的海马神经回路的活动和连接性 用于与 AW 相关的癫痫发作。在目标 1 中，我们将确定 AW 是否会改变神经元和功能连接 通过使用狂犬病病毒和小麦胚芽凝集素 (WGA) 介导的逆行和顺行追踪来检测 DGC 方法，分别。我们还将使用多个 DREADD 并评估 de novo 神经网络的重要作用 在 AW 相关癫痫发作中，海马新生儿 DGC 和输入神经元之间形成的回路。目标2，使用 Ca2+ 成像和各种磁共振成像 (MRI) 模式使我们能够进行纵向研究， 我们将确定与 AW 相关的海马和整体神经回路的活动和连接性 癫痫发作。在目标 3 中，我们将识别并验证可能导致突触和神经元改变的转录组 海马新生 DGC 响应 AE 和 AW 的连接。单细胞RNA测序的用途 将使我们不仅能够以细胞类型特异性的方式记录差异表达的基因，而且还可以确定 在 AE 和 AW 期间区分病理性新生儿 DGC 和正常 DGC 的转录组。共， 我们的提案将剖析新生儿海马的分子、细胞和神经回路机制 DGC 是 AW 相关癫痫发作的基础。