The role of sex- and cell-type specific protein degradation increases in the amygdala in fear memory formation

杏仁核中性别和细胞类型特异性蛋白质降解在恐惧记忆形成中的作用增加

基本信息

批准号：
10117396
负责人：
TIMOTHY JOSEPH JAROME
金额：
$ 41.67万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2025-10-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10117396
关键词：
Affect Amygdaloid structure Behavioral Model Binding Biological Biological Assay Brain Brain region Cell Nucleus Cells Clustered Regularly Interspaced Short Palindromic Repeats Code DNA DNA Methylation DNMT3a Development Disease Emotional Etiology Female Fright Gene Expression Genes Genetic Transcription Germ Cells Goals Human Impairment Knowledge Lead Learning Literature Mass Spectrum Analysis Mediating Memory Memory Disorders Mental disorders Methylation Modeling Molecular Nuclear Nuclear Protein Pharmacology Physiological Polyubiquitination Population Post-Traumatic Stress Disorders Predisposition Process Promoter Regions Protein Biosynthesis Proteins Rattus Regulation Rodent Role Sex Differences Shock Symptoms System Technology Testing Training Transcriptional Regulation Translational Research Ubiquitin Ubiquitination Work base cell type conditioned fear design experimental study fear memory in vivo male memory retention multicatalytic endopeptidase complex next generation novel therapeutic intervention protein degradation response sex therapeutic development transcriptome sequencing ubiquitin mediated proteasome degradation whole genome

Affect Amygdaloid structure Behavioral Model Binding Biological Biological Assay Brain Brain region Cell Nucleus Cells Clustered Regularly Interspaced Short Palindromic Repeats Code DNA DNA Methylation DNMT3a Development Disease Emotional Etiology Female Fright Gene Expression Genes Genetic Transcription Germ Cells Goals Human Impairment Knowledge Lead Learning Literature Mass Spectrum Analysis Mediating Memory Memory Disorders Mental disorders Methylation Modeling Molecular Nuclear Nuclear Protein Pharmacology Physiological Polyubiquitination Population Post-Traumatic Stress Disorders Predisposition Process Promoter Regions Protein Biosynthesis Proteins Rattus Regulation Rodent Role Sex Differences Shock Symptoms System Technology Testing Training Transcriptional Regulation Translational Research Ubiquitin Ubiquitination Work base cell type conditioned fear design experimental study fear memory in vivo male memory retention multicatalytic endopeptidase complex next generation novel therapeutic intervention protein degradation response sex therapeutic development transcriptome sequencing ubiquitin mediated proteasome degradation whole genome

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项目摘要

Project Summary/Abstract The broad goal of this proposal is to understand how fear memories are formed and stored in the brain. Post- traumatic stress disorder (PTSD) affects nearly 5% of the world population, however, current treatments have limited efficacy in reversing the symptoms of this disorder. Furthermore, females are more likely than males to develop PTSD, though the mechanisms controlling this sex-dependent predisposition remain equivocal. Recently, work from our group and others have implicated protein degradation mediated by the ubiquitin- proteasome system (UPS) in fear memory formation in the amygdala, the primary brain region involved in emotional processing. However, the functional role of protein degradation in fear memory formation has yet to be identified. Furthermore, previous studies on UPS-mediated protein degradation have focused exclusively on males, so little is known about whether similar UPS mechanisms control the formation of fear memories in females. In our preliminary studies, we found that in males protein degradation was localized to the nucleus of amygdala cells following fear learning, suggesting a potential role in transcriptional control, though this has never been directed tested. Surprisingly, females did not show any changes in UPS-mediated protein degradation in the amygdala following fear learning though did have elevated baseline ubiquitin-proteasome activity in comparison with males, which was associated with increased DNA 5-hydroxymethylation (5-hmc) of Uba52, one of the ubiquitin coding genes. This suggests that elevated baseline UPS activity in females could be regulated by altered DNA methylation of Uba52 and may be sufficient for them to acquire fear memories. The work in this proposal is designed to answer these important questions about sex-dependent differences in the role of nuclear protein degradation in transcriptional control during fear memory formation. Using a combination of pharmacology and sophisticated CRISPR-dCas9 manipulations of proteasome activity in combination with mass spectrometry and whole genome next generation RNA-seq technology, Aim 1 will test whether nuclear protein degradation is involved in transcriptional control during fear memory formation and whether this varies between males and females. Aim 2 will use cutting-edge CRISPR-dCas9 technology to control the methylation state of Uba52 and will test how this effects baseline differences in ubiquitin- proteasome activity in the amygdala of males and females. Finally, in Aim 3 we will using CRISPR-dCas9 technology to test whether increased baseline protein degradation in the amygdala of females primes them to acquire fear memories. Collectively, this study will answer important questions about sex-dependent differences in the role of protein degradation in fear memory formation. The results obtained from this project could have important implications for understanding the etiology of sex-related differences in fear memory formation and lead to the development of novel therapeutic strategies to treat major psychiatric disorders.

项目摘要/摘要该提议的广泛目标是了解如何形成和存储大脑中的恐惧记忆。邮政- 创伤应激障碍（PTSD）影响了近5％的世界人口，但是，当前的治疗方法具有逆转这种疾病症状的功效有限。此外，女性比男性更有可能发展为PTSD，尽管控制这种性别依赖性倾向的机制仍然是模棱两可的。最近，我们小组和其他人的工作暗示了蛋白质降解由泛素 - 杏仁核的恐惧记忆形成中的蛋白酶体系统（UPS），涉及的主要脑区域情绪处理。但是，蛋白质降解在恐惧记忆形成中的功能作用尚未被识别。此外，先前关于UPS介导的蛋白质降解的研究仅集中在男性，关于类似的UPS机制是否控制着恐惧记忆的形成知之甚少女性。在我们的初步研究中，我们发现在雄性中，蛋白质降解位于害怕学习后的杏仁核细胞，表明在转录控制中可能作用，尽管这有从未被指导测试。令人惊讶的是，女性没有显示UPS介导的蛋白质的任何变化害怕学习后，杏仁核的退化虽然基线泛素 - 蛋白酶体确实升高与男性相比，与男性的活性与增加的DNA 5-羟基甲基化（5-HMC）相比 UBA52，泛素编码基因之一。这表明女性的基线UPS活动升高可能受UBA52的DNA甲基化改变的调节，可能足以使他们获得恐惧记忆。该提案的工作旨在回答有关性别依赖性差异的这些重要问题核蛋白降解在恐惧记忆形成期间转录控制中的作用。使用药理学和复杂的CRISPR-DCAS9操纵蛋白酶体活动的结合结合质谱和整个基因组下一代RNA-seq技术，AIM 1将测试核蛋白降解是否参与恐惧记忆形成期间的转录控制和这是否在男性和女性之间有所不同。 AIM 2将使用尖端的CRISPR-DCAS9技术来控制UBA52的甲基化状态，并将测试这种影响泛素的基线差异男性和女性杏仁核中的蛋白酶体活性。最后，在AIM 3中，我们将使用CRISPR-DCAS9 测试女性杏仁核中基线蛋白质降解是否增加的技术是否会质疑它们获得恐惧的回忆。总的来说，这项研究将回答有关性依赖性的重要问题蛋白质降解在恐惧记忆形成中的作用差异。从这个项目获得的结果对于理解与性别相关的性别差异的病因可能具有重要意义形成并导致发展新型治疗策略来治疗主要的精神疾病。