Linking stress-associated brain and adipose functions

项目摘要/摘要 急性高强度应激通过脑底物影响能量稳态的基本机制 人们对这些过程的性别二态知之甚少。环境应激源引发争斗-或- 通过脑干蓝斑(LC)的飞行或冻结反应，蓝斑是一种主要的交感神经调节物质。 触发能量新陈代谢的人和动物。LC支配棕色脂肪组织(BAT)和 高水平表达神经肽PACAP(脑垂体腺苷环化酶激活)的PAC1受体 多肽)。PACAP/PAC1是各种应激源的关键调节者，包括恐惧、食欲、进食和能量 新陈代谢，并在基因上与人类创伤后应激障碍(PTSD)等应激相关疾病有关。 有趣的是，患有创伤后应激障碍的女性血液中这种神经肽和脑PAC1受体水平很高 在动物身上以一种性别分化的方式调节恐惧。检测PAC1在急性重症患者中的重要性 应激和能量平衡，我们在小鼠中使用了应激增强恐惧学习(SEFL)行为模型。 我们的初步数据显示，从LC中去除PAC1受体可以增强SEFL的表达和能量 通过间接量热法评估代谢，并增加新陈代谢基因，如解偶联 雌性蝙蝠中蛋白质1的含量高于雄性蝙蝠。因此，利用PAC1受体的生物学基础，我们的目标是建立一个 在系统和性别依赖的情况下，连接急性强烈应激和能量代谢的机制框架 方式通过LC。我们假设LC中的PAC1受体在门控应激相关中起重要作用 新陈代谢信息和BAT以性别二态的方式发挥作用。目标1将测试SEFL中介的损失 研究LC-PAC1对全身能量消耗和BAT功能的影响。为此，我们将测试是否存在病毒 Cre-重组酶介导的PAC1缺失对Sefl相关基因表达的影响 蝙蝠的能量消耗和产热基因的诱导与性别相关(通过转录本 分析、线粒体生物能量学和组织脂肪分解)。Aim 2将测试PAC1是否表达LC神经元 在SEF1的领导下，直接投射到蝙蝠，以性二态的方式。为此，我们将使用逆行病毒跟踪 结合PAC1的原位杂交，确定LC-BAT投射细胞群。我们 预测雌性小鼠将表现出能量消耗增加，BAT产热和LC增加 投射到击球。总体而言，使用最先进的工具和技术以及严格的系统生物学方法 将急性紧张应激与能量代谢联系起来，我们的研究将为未来的几个 工作。这将成为研究慢性应激时新陈代谢的中枢交感神经控制的前提。 相关的疾病，如创伤后应激障碍，与代谢性疾病并存，并在 美国人口。

PROJECT SUMMARY/ABSTRACT The fundamental mechanisms by which acute intense stress affects energy homeostasis via brain substrates and sexual dimorphism of these processes are poorly understood. Environmental stressors induce fight-or- flight-or-freeze response via the locus coeruleus (LC) in the brainstem, a major sympathetic regulator in humans and animals that triggers energy metabolism. LC innervates the brown adipose tissue (BAT) and expresses high levels of PAC1 receptor for the neuropeptide PACAP (pituitary adenylate cyclase activating peptide). PACAP/PAC1 are critical regulators of stressors of all kinds, fear, appetite, feeding, and energy metabolism and genetically linked to stress-related disorders like post-traumatic stress (PTSD) in humans. Intriguingly, women with PTSD showing high blood levels of this neuropeptide and brain PAC1 receptors regulate fear in a sexually dimorphic manner in animals. To test the importance of PAC1 in acute intense stress and energy homeostasis, we used the stress-enhanced fear learning (SEFL) behavioral model in mice. Our preliminary data show that ablation of PAC1 receptors from the LC enhances SEFL expression, energy metabolism as assessed by indirect calorimetry measurement, and increases metabolic genes like uncoupling protein 1 in BAT in females than males. Thus, harnessing on the biology of PAC1 receptors, we aim to lay a mechanistic framework linking acute intense stress and energy metabolism in a systematic and sex-dependent manner via the LC. We hypothesize that PAC1 receptors in the LC are important for gating stress-associated metabolic information and BAT functions in a sexually dimorphic manner. Aim 1 will test a SEFL mediated loss of function of LC-PAC1 on whole body energy expenditure and on BAT functions. For this, we will test if viral cre-recombinase mediated PAC1 deletion from LC in mice with floxed PAC1 regulates SEFL-associated energy expenditure and induction of thermogenic genes in BAT in a sex-dependent manner (via transcript analysis, mitochondrial bioenergetics, and tissue lipolysis). Aim 2 will test if PAC1 expressing LC neurons project directly to BAT in a sexually dimorphic manner under SEFL. For this, we will use retrograde viral tracing to determine LC to BAT projecting cell populations combined with in situ hybridization for PAC1 in LC. We predict that female mice will show enhanced energy expenditure, BAT thermogenesis and increased LC projection to BAT. Overall, using state-of-the-art tools and techniques and rigorous systems biology approach of linking acute intense stress and energy metabolism, our studies will lay crucial groundwork for several future work. This will serve as a premise for studying central sympathetic control of metabolism in chronic stress- related conditions such as PTSD that are comorbid with metabolic diseases and increasingly prevalent in the US populations.