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

• 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
• 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

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 这项研究的主要目标是了解恐惧记忆是如何在大脑中形成和储存的。邮寄- 创伤应激障碍(PTSD)影响着近5%的世界人口，然而，目前的治疗方法已经 在扭转这种疾病症状方面的效果有限。此外，女性比男性更有可能 发展为创伤后应激障碍，尽管控制这种性别依赖性倾向的机制仍然不明确。 最近，我们团队和其他人的研究表明，泛素介导的蛋白质降解- 蛋白酶体系统(UPS)在杏仁核恐惧记忆形成中的作用 情绪处理。然而，蛋白质降解在恐惧记忆形成中的功能作用尚未得到证实 被指认出来。此外，以前对UPS介导的蛋白质降解的研究仅集中在 男性，所以关于类似的UPS机制是否控制着男性恐惧记忆的形成，人们知之甚少 女性。在我们的初步研究中，我们发现在男性中，蛋白质降解定位于 杏仁核细胞跟随恐惧学习，暗示在转录控制中可能发挥作用，尽管这 从未被直接测试过。令人惊讶的是，女性在UPS介导的蛋白质中没有表现出任何变化 尽管在恐惧学习后杏仁核的退化确实有基线泛素蛋白酶体的升高 与男性相比，这与DNA5-羟甲基化(5-HMC)增加有关 Uba52，泛素编码基因之一。这表明，女性体内基线UPS活性升高可能 受Uba52基因甲基化改变的调控，可能足以让他们获得恐惧记忆。 这项提案中的工作旨在回答关于性别依赖差异的这些重要问题 核蛋白降解在恐惧记忆形成过程中转录调控中的作用。使用 药理学和复杂的CRISPR-dCas9对蛋白酶体活性的调控相结合 结合质谱学和全基因组下一代rna-seq技术，Aim 1将进行测试 核蛋白降解是否参与恐惧记忆形成和转录控制 这一点在男性和女性之间是否存在差异。AIM 2将使用尖端CRISPR-dCas9技术来 控制Uba52的甲基化状态，并将测试这如何影响泛素的基线差异 雄性和雌性杏仁核中的蛋白酶体活性。最后，在目标3中，我们将使用CRISPR-dCas9 测试女性杏仁核基线蛋白降解增加是否会使她们 获得恐惧记忆。总而言之，这项研究将回答有关性依赖的重要问题 不同的蛋白质降解在恐惧记忆形成中的作用。从这个项目中获得的成果 可能对理解与性别有关的恐惧记忆差异的原因有重要意义 形成并导致开发新的治疗策略来治疗严重的精神疾病。