TNF-α Signaling in Noise Trauma-Induced PV Neuron Loss and Dysfunction

噪声创伤引起的 PV 神经元丢失和功能障碍中的 TNF-α 信号转导

• 批准号: 10664648
• 负责人: Shaowen BAO
• 金额: $ 23.03万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10664648
• 关键词: Acceleration; Acute; Auditory; Auditory Perceptual Disorders; Auditory area; Behavior; Brain; Brain Diseases; Cell Death; Cell Survival; Cells; Custom; Cytoprotection; Data; Detection; Disease; Enterobacteria phage P1 Cre recombinase; Exposure to; Functional disorder; Image; Immune response; Inflammatory; Interneurons; Link; Mediating; Mus; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Neurotransmitters; Noise; Optics; Output; Parvalbumins; Pathology; Phenotype; Phosphorylation; Physiologic pulse; Population; Probability; Publishing; RIPK3 gene; Receptors, Tumor Necrosis Factor, Type II; Reporter; Research; Rodent Model; Role; Signal Transduction; Slice; Synapses; TNF gene; TNFRSF1A gene; TNFRSF1B gene; Testing; Tinnitus; Transfection; Traumatic Brain Injury; Tumor Necrosis Factor Receptor; Viral; Viral Vector; Visualization; auditory pathway; auditory processing; cytokine; design; hippocampal pyramidal neuron; knock-down; mouse model; nervous system disorder; neuroinflammation; neuron loss; noise exposure; noise trauma; optogenetics; patch clamp; prevent; receptor; response; small hairpin RNA; synaptic inhibition

PROJECT SUMMARY/ABSTRACT Noise trauma can lead to loss of parvalbumin-positive inhibitory interneurons in the auditory cortex, which is associated with audiotory processing deficit and tinnitus in rodent models. The mechanisms underlying noise- induced PV neuron loss are unknown. We propose to examine the hypothesis that differential activation of TNFR1 and TNFR2 in cortical PV neurons determines the fate of the PV neurons following noise trauma, with TNFR1 biasing for, and TNFR2 biasing against, neuronal loss and dysfunction. Specific Aim 1. Determine the effects of TNFR1 or TNFR2 knockdown on noise-induced PV neuron loss. PV-Cre-tdTomato mice will be injected with one of three viral vectors (with TNFR1 shRNA, TNFR2 shRNA or scrambled sequences as a control) in the auditory cortex, and exposed or sham-exposed to noise trauma. PV neurons will be visualized by the Cre reporter tdTomato in auditory cortical sections. Transfected neurons will be visualized with the viral reporter GFP. Our hypothesis predicts that noise-induced PV neuron loss will be reduced by TNFR1 knockdown, but enhanced by TNFR2 knockdown for the transfected PV neurons. Cell loss should not be altered for the populations of un-transfected PV neurons, and PV neurons transfected with the scrambled sequences. Specific Aim 2. Examine the effects of TNFR1 or TNFR2 knockdown on noise-induced dysfunction of PV neuron synapses. Our pilot data indicate that noise trauma leads to a reduced transmitter release probability at the output synapses of the PV neurons, and accelerated neurotransmitter depletion. We hypothesize that this synaptic dysfunction depends on the activation of TNFR1 in the surviving PV neurons, and knockdown of TNFR1 will prevent the synaptic deficits. In addition, knockdown of TNFR2 should exacerbate PV neuron dysfunction. PV-Cre-ChR2-tdTomato mice will be injected with one of the three viral vectors in the auditory cortex, and be exposed or sham-exposed to noise trauma. We will record optically activated inhibitory synaptic current in Layer2/3 pyramidal neurons in acute auditory cortical slices. Synaptic input-output curve, paired-pulse modulation and depletion will be examined. Afterward, the slices will be fixed and imaged to quantify PV neuron loss and viral transfection rate in the surviving PV neurons, which will then be correlated with PV neuron synaptic dysfunction.We hypothesize that noise exposure disrupts cortical PV neuron function, and PV neuron dysfunction is a cause of gap detection deficit. We propose to use a mouse model to test this central hypothesis in the follwing specific aims.

项目摘要/摘要 噪声创伤会导致听觉皮层中的白蛋白阳性抑制性抑制性中间神经元的丧失，这是 与啮齿动物模型中的唱机处理不足和耳鸣相关。噪声的基础机制 - 诱导的PV神经元丧失尚不清楚。我们提出了以下假设 皮质PV神经元中的TNFR1和TNFR2决定了噪声创伤后PV神经元的命运， TNFR1偏向于偏置，偏向神经元丧失和功能障碍。 具体目标1。确定TNFR1或TNFR2敲低对噪声诱导的PV神经元丧失的影响。 PV-CRE-TDTOMATO小鼠将注射三个病毒载体之一（用TNFR1 shRNA，TNFR2 shRNA或 在听觉皮层中，炒序列是对照），并暴露于噪声创伤。 PV 在听觉皮质部分中，CRE记者TDTOMATO将可视化神经元。转染的神经元会 用病毒记者GFP可视化。我们的假设预测，噪声引起的PV神经元丧失将是 通过TNFR1敲低减少，但转染的PV神经元的TNFR2敲低增强。细胞损失 未转染的PV神经元的种群不应改变 炒序列。 特定目标2。检查TNFR1或TNFR2敲低对噪声诱导的PV功能障碍的影响 神经元突触。我们的飞行员数据表明​​，噪声创伤导致发射机释放概率降低 在PV神经元的输出突触中，并加速神经递质耗竭。我们假设这是 突触功能障碍取决于幸存的PV神经元中TNFR1的激活，而TNFR1敲低 将防止突触缺陷。此外，TNFR2的敲低应加剧PV神经元功能障碍。 PV-CRE-CHR2-TDTOMATO小鼠将在听觉皮层中注射三个病毒载体之一，并为 暴露于噪声创伤或假性暴露。我们将记录光学激活的抑制性突触电流 急性听觉皮质切片中的层/3锥体神经元。突触输入输出曲线，配对脉冲 将检查调节和耗竭。之后，切片将被固定并成像以量化PV神经元 存活的PV神经元中的损失和病毒转染率，然后将与PV神经元突触相关 功能障碍。我们假设噪声暴露会破坏皮质PV神经元功能和PV神经元 功能障碍是差距检测缺陷的原因。我们建议使用鼠标模型测试此中心假设 在特定目标中。