Understanding Foxp2-Mediated Molecular Signaling in Fear Learning

项目摘要/摘要 创伤后应激障碍(PTSD)是一种常见的、使人衰弱的精神疾病，可利用的药物有限 个性化治疗。最近的全基因组联合研究(GWAS)已经开始阐明 与这种疾病有关的重要靶基因。最近发现的这些靶标之一就是转录 Foxp2因子。Foxp2此前曾被认为与语言学习障碍有关，然而，它在成人中的作用 恐惧学习是未知的。在成年小鼠中，Foxp2 mRNA在周围的插入细胞团中表达 杏仁基底外侧核。间质细胞对条件性恐惧反应和恐惧具有重要的功能 然而，Foxp2转录因子、其上游介体和下游靶标的作用， 杏仁核中与恐惧相关的行为在调节中的作用尚不清楚。最近的证据表明，Foxp2 受Wnt信号通路的多个组成部分调节，进而在反馈环中作用于 调节Wnt途径成员的调节。在这个提案中，我将测试Foxp2调控的假设 恐惧学习的特定成分通过其与Wnt信号通路的相互作用 杏仁核的细胞。在目标1中，我将测试恐惧时杏仁核中Foxp2的转录调控 学习。我将使用RT-PCR对恐惧学习后的Foxp2转录进行时程分析 RNAScope。我还将在Foxp2基因敲除后使用单核测序来表征Foxp2靶标 恐惧学习后杏仁核内特定的细胞类型。最后，我将执行与恐惧相关的行为 雄性和雌性成年小鼠杏仁核特异性Foxp2基因敲除的特征。在AIM2中，我将 机械地检验Foxp2在恐惧学习过程中受Wnt信号通路调控的假设， 建立在Ressler实验室之前的工作基础上，该工作表明Wnt信号通路组件在成人中的作用 学习和记忆。概述的K08提案将建立在我之前在发展道路上的经验基础上， 分子生物学和遗传学，同时在疾病相关神经的哺乳动物模型中继续训练 电路。这项建议的最终目的是在收集的同时提高我的技术和专业技能 数据将构成在成为独立公司的道路上获得独立资金的基础 医生-科学家。

Project Summary/Abstract Post-Traumatic Stress Disorder (PTSD) is a common and debilitating mental illness with limited available personalized treatments. Recent Genome-Wide Association Studies (GWAS) have begun to elucidate significant target genes implicated in this disease. One of these recently identified targets is the transcription factor Foxp2. Foxp2 has been previously implicated in language learning disorders, however, its role in adult fear learning is unknown. In adult mice, Foxp2 mRNA is expressed in clusters of Intercalated Cells surrounding the basolateral amygdala. Intercalated Cells are functionally important for conditioned fear response and fear extinction; however, the role of the Foxp2 transcription factor, its upstream mediators and downstream targets, in modulating fear-related behaviors in the amygdala remains unknown. Recent evidence suggests that Foxp2 is regulated by multiple components of the Wnt signaling pathway and, in turn, acts in a feedback loop to modulate regulation of Wnt pathway members. In this proposal, I will test the hypothesis that Foxp2 regulates specific components of fear learning through its interaction with the Wnt signaling pathway in the Intercalated Cells of the amygdala. In Aim 1, I will test transcriptional regulation of Foxp2 in the amygdala during fear learning. I will perform a time-course analysis of Foxp2 transcription after fear learning using RT-PCR and RNAscope. I will also use single nuclei sequencing following Foxp2 knockout to characterize Foxp2 targets in specific cell types within amygdala after fear learning. Finally, I will perform fear-related behavioral characterization with amygdala-specific Foxp2 knockout using male and female adult mice. In Aim2, I will mechanistically test the hypothesis that Foxp2 is regulated by Wnt signaling pathway during fear learning, building on prior work from the Ressler Lab that showed the role of Wnt signaling pathway components in adult learning and memory. The outlined K08 proposal will build on my prior experience in developmental pathways, molecular biology and genetics, while continuing training in a mammalian model of disease-relevant neural circuitry. The ultimate goal of this proposal is to further my technical and professional skills while collecting data that will form the basis for obtaining independent funding on a path to becoming an independent Physician-Scientist.