Long non-coding enhancer RNA controls stress-induced changes in reward-related behaviors

项目总结/摘要 慢性压力改变了奖励相关的行为，这伴随着神经系统的变化。 回路功能，特别是涉及内侧前额叶皮层（mPFC）。但 控制回路功能障碍的分子机制及其对相关行为的影响 仍然知之甚少。 Npas 4基因编码神经元PAS结构域蛋白4（NPAS 4）转录因子 作为兴奋性和抑制性突触连接的稳态调节器， 对突触活动变化的反应。我们在老鼠身上的初步发现表明， 应激经历触发了mPFC中NPAS 4表达的瞬时诱导， 慢性社会失败压力（CSDS）诱导的奖励相关行为缺陷。 此外，这些研究表明，NPAS 4是CSD诱导的mPFC减少所必需的 锥体神经元兴奋性传递和树突棘密度。 有趣的是，虽然蛋白质编码基因（mRNA）对功能性 调节大脑活动，人类和啮齿动物的基因组编码大量的长 非编码RNA（lncRNA）。新出现的证据表明lncRNA在 基因表达，但单个lncRNA的生理和病理功能仅 开始被探索。为了解决这一知识缺口，我们将研究一种新颖的，无注释的， lnc-增强子RNA（lnc-eRNA）从Npas 4（Npas 4 eRNA）的增强子区域转录。我们 发现Npas 4 eRNA是Npas 4 mRNA表达和CSDS诱导的缺陷所必需的。 蔗糖偏好Npas 4 eRNA形成DNA：RNA杂合R环结构， Npas 4 mRNA表达。我们假设Npas 4 eRNA及其R环结构直接连接 增强子相关的转录机制与Npas 4转录起始位点通过3D 染色质环结构。 在这项提案中，我们将联合收割机结合创新的分子和细胞方法， 男性和女性的心理社会和心理应激源的行为学相关模型 研究Npas 4 eRNA和R-loop在应激诱导的大鼠PFC中的功能。 表观遗传基因调控，功能和形态突触可塑性，以及 与奖励有关的行为。

Project Summary/Abstract Chronic stress alters reward-related behavior and this is accompanied by changes in neural circuit function, especially that involving the medial prefrontal cortex (mPFC). However, the molecular mechanisms controlling circuit dysfunction and its effect on associated behaviors remain poorly understood. The Npas4 gene encodes the Neuronal PAS domain protein 4 (NPAS4) transcription factor that functions as a homeostatic regulator of excitatory and inhibitory synaptic connections in response to changes in synaptic activity. Our preliminary findings in mice show that aversive stress experience trigger the transient induction of NPAS4 expression in mPFC and that this is required for chronic social defeat stress (CSDS)-induced reward-related behavior deficit. Furthermore, these studies show that NPAS4 is required for CSDS-induced reduction of mPFC pyramidal neuron excitatory transmission and dendritic spine density. Interestingly, while protein-coding genes (mRNA) are obviously important for functional regulation of brain activity, human and rodent genomes encode a much larger number of long non-coding RNAs (lncRNAs). Emerging evidence demonstrates significant roles for lncRNAs in gene expression, but the physiological and pathological functions of individual lncRNAs are only beginning to be explored. To address this knowledge gap, we will study a novel, non-annotated, lnc-enhancer RNA (lnc-eRNA) transcribed from the enhancer region of Npas4 (Npas4eRNA). We found that Npas4eRNA is required for Npas4mRNA expression and for CSDS-induced deficits in sucrose preference. Npas4eRNA forms an DNA:RNA hybrid R-loop structure that is required for Npas4mRNA expression. We hypothesize that the Npas4eRNA and its R-loop structure directly link the enhancer-associated transcriptional machinery with the Npas4 transcription start site via 3D chromatin loop structure. In this proposal, we will combine innovative molecular and cellular approaches with ethologically relevant models of psychosocial and psychological stressors in both male and female mice and examine the function of Npas4eRNA and R-loop in the PFC for the stress-induced epigenetic gene regulation, functional and morphological synaptic plasticity, and alterations in reward-related behaviors.