The Circadian Molecular Clock is a Biomarker for Epilepsy in Focal Cortical Dysplasia

昼夜节律分子钟是局灶性皮质发育不良中癫痫的生物标志物

• 批准号: 10351603
• 负责人: Judy Shih-Hwa Liu
• 金额: $ 7.72万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10351603
• 关键词: Administrative Supplement; Bioinformatics; Biological Markers; Brain region; Chronic; Cortical Dysplasia; Development; Epilepsy; Excision; Excitatory Synapse; Hippocampus (Brain); Histological Techniques; Human; Inhibitory Synapse; Knockout Mice; Laboratories; Learning; Maintenance; Mediating; Microscopy; Molecular Analysis; Mus; Neurologic; Neurons; Neurotransmitters; Partial Epilepsies; Presynaptic Terminals; Proteins; Publishing; RNA analysis; Regulation; Reporting; Role; Seizures; Stains; Synapses; System; Techniques; Testing; Therapeutic; Tissues; Training; bZIP Protein; base; brain tissue; circadian; excitatory neuron; experience; hippocampal pyramidal neuron; inhibitory neuron; molecular clock; mutant; nano-string; synaptogenesis; transcription factor; transcriptome; transcriptome sequencing

Abstract Focal epilepsy is the most common cause of epilepsy. The abnormal epileptogenic region of the brain that generates seizures or "seizure focus" is critically important because its removal often results in a complete cure of the epilepsy. However, this tissue is under studied. Our first transcriptome study demonstrated diminished expression of the CLOCK transcription factor in seizure focus in contrast to control tissue. In addition, the downstream targets of CLOCK, the PAR bZip transcriptions factors are also diminished in the seizure focus. The deletion of Par bZip transcription factors in mice causes epilepsy. Thus, the role of CLOCK in the seizure focus may be through its regulation of the Par bZip transcription factors. In our study of CLOCK, we find that the deletion of CLOCK changes synaptic numbers, especially in inhibitory synapses. However, the function of PAR bZip factors per se in synaptogenesis and synaptic maintenance has not been explored, despite reports of their roles in regulating neurotransmitter levels. Based on our published finding that increased neuronal hyperexcitability of CLOCK-deficient mutants is associated with changes in the number of inhibitory synaptic terminals on excitatory pyramidal neurons, we hypothesize that these changes are mediated by PAR bZip. The Aims of this Supplement will test the hypothesis that, as downstream effectors of CLOCK, PAR bZip transcription factors govern synaptic maintenance and stability (Aim 1), as well as the expression levels of synaptic proteins (Aim 2). Both Aims will utilize the triple PAR bZip knockout mouse, so that Sean will gain experience in histological techniques and advanced microscopy, as well as molecular analyses and bioinformatics.

抽象的 局灶性癫痫是癫痫最常见的原因。异常致痫 产生癫痫或“癫痫焦点”的大脑区域至关重要，因为它 去除通常可以完全治愈癫痫。然而，这种组织仍在研究中。 我们的第一个转录组研究表明 CLOCK 转录的表达减少 与对照组织相比，癫痫病灶的因素。此外，下游目标 CLOCK、PAR bZip 转录因子在癫痫病灶中也减少。这 小鼠中 Par bZip 转录因子的缺失会导致癫痫。因此，CLOCK 的作用 癫痫发作的焦点可能是通过其对 Par bZip 转录因子的调节。 在我们对 CLOCK 的研究中，我们发现 CLOCK 的删除会改变突触数量， 特别是在抑制性突触中。然而，PAR bZip 的功能本身就影响了 尽管有关于突触发生和突触维持作用的报道，但尚未对其进行探索 调节神经递质水平。根据我们发表的发现，增加了神经元 CLOCK缺陷突变体的过度兴奋性与细胞数量的变化有关 兴奋性锥体神经元上的抑制性突触末端，我们假设这些 变化是由 PAR bZip 介导的。本补充文件的目的将检验以下假设： 作为 CLOCK 的下游效应器，PAR bZip 转录因子控制突触 维持和稳定性（目标 1），以及突触蛋白的表达水平（目标 2）。 两个 Aims 都将使用三重 PAR bZip 敲除鼠标，以便 Sean 获得以下方面的经验： 组织学技术和先进的显微镜，以及分子分析和 生物信息学。