Postmortem studies of CRF-PACAP in human PTSD (Berretta)

摘要：项目5(人类创伤后应激障碍/贝雷塔的PACAP-CRF的尸检研究) 令人信服的证据表明，促肾上腺皮质激素释放因子(CRF)和垂体腺苷酸环化酶激活 多肽(PACAP)及其相互作用在应激反应中起关键作用， 焦虑、昼夜节律调节与创伤后应激障碍的病理生理机制 (创伤后应激障碍)。人们对涉及的潜在神经回路知之甚少，但重要的线索指向床 终纹核(BNST)、杏仁核(AMG)、扣带回背侧前核(DACG)和 下丘脑(HPTH)作为这些功能的关键调节。目前的证据表明，CRF和PACAP 导致创伤后应激障碍的机制可能是性别特有的，这增加了潜在大脑 男性和女性的变化和潜在的治疗靶点可能不同。目前和初步的数据表明 PACAP信号可能直接影响CRF表达细胞和PACAP及其基因的昼夜表达 同源受体PAC1R及其随后对CRF系统的调节在发病过程中可能不同。 天。这种变化可能会潜在地导致与DSM定义的睡眠/唤醒周期相关的中断 疾病包括创伤后应激障碍、严重抑郁症和焦虑症。令人惊讶的是，几乎没有任何信息是 可在细胞水平表达CRF和PACAP信号通路在人AMG、BNST、DACG和 HPTH，它们与昼夜节律的关系，以及CRF/PACAP相互作用在 创伤后应激障碍的神经病理学。我们的主要假设是影响CRF/PACAP通路的异常在 BNST、AMG、DACG和HPTH参与了PTSD的病理过程。在目标1中，我们解决了一个关键差距 了解CRF、PACAP和它们的区域、性别和昼夜节律特异性的表达和分布 健康人脑中的受体。我们的假设是CRF、PACAP和 它们的受体是区域和性别特异性的；特别是，我们预测PACAP受体将显示细胞-和 CRF阳性神经元的性别特异性和表达，支持PACAP调节这些神经元的假说 神经元以性别依赖的方式。在目标2和目标3中，我们检查是否-在蛋白质、基因表达， PTSD患者DACG、AMG、BNST和HPTH的细胞水平信号通路发生改变。我们的假设 与健康对照组相比，创伤后应激障碍患者的CRF和PACAP信号通路将发生改变， 随着PACAP表达的增加与CRF信号通路的变化相关，在一个地区--性别--和 昼夜节律特有的方式。我们预测PACAP阳性细胞和轴突的增加，反映了 局部和下丘脑输入的PACAP表达增加将伴随着表达的改变 在CRF阳性细胞中PACAP受体和下游信号通路的表达。项目5可确定CRF和 PACAP系统和回路在应激相关疾病(如创伤后应激障碍)中发生根本性改变，是一种 加强其他(临床前、临床)要素并由其增强的中心的关键关系。

SUMMARY: PROJECT 5 (POSTMORTEM STUDIES OF PACAP-CRF IN HUMAN PTSD/BERRETTA) Compelling evidence indicates that corticotropic releasing factor (CRF) and pituitary adenylyl cyclase-activating polypeptide (PACAP), as well as their interactions together, make critical contributions to stress responses, anxiety, circadian rhythm regulation, and the pathophysiological mechanisms of post-traumatic stress disorder (PTSD). The underlying neural circuitry involved is poorly understood, but important clues point to the bed nucleus of the stria terminalis (BNST), amygdala (AMG), dorsal anterior cingulate gyrus (dACG), and the hypothalamus (HPTh) as critical regulators of these functions. Current evidence indicates that CRF and PACAP mechanisms that contribute to PTSD may be sex-specific, raising the possibility that the underlying brain changes and potential therapeutic targets may differ in males and females. Current and preliminary data suggest that PACAP signaling may directly affect CRF expressing cells and that circadian expression of PACAP and its cognate receptor PAC1R, and their subsequent regulation of CRF systems, may vary during the course of the day. Such variations may potentially contribute to disruptions of sleep/wake cycles associated with DSM-defined illnesses including PTSD, major depression, and anxiety disorders. Surprisingly, virtually no information is available on cell-level expression of CRF and PACAP signaling pathways in the human AMG, BNST, dACG and HPTh, their relationships to circadian rhythms, and the involvement of CRF/PACAP interactions in the neuropathology of PTSD. Our overarching hypothesis is that abnormalities affecting CRF/PACAP pathways in the BNST, AMG and dACG and HPTh contribute to the pathology of PTSD. In Aim 1, we address a critical gap of knowledge on the region-, sex- and circadian- specific expression and distribution of CRF, PACAP, and their receptors in healthy human brain. Our hypothesis is that protein and mRNA expression of CRF, PACAP, and their receptors are region- and sex-specific; in particular, we predict that PACAP receptors will show cell- and sex- specificity and expression in CRF-positive neurons, supporting the hypothesis that PACAP regulates these neurons in a sex dependent manner. In Aims 2 and 3, we examine whether—at the protein, gene expression, and cellular level—signaling pathways in the dACG, AMG, BNST and HPTh are altered in PTSD. Our hypothesis is that CRF and PACAP signaling pathways will be altered in subjects with PTSD, relative to healthy controls, with increased PACAP expression correlated with CRF signaling pathway changes, in a region-, sex-, and circadian rhythm-specific manner. We predict that increases of PACAP-positive cells and axons, reflecting increased PACAP expression locally and from hypothalamic inputs, will be accompanied by altered expression of PACAP receptors and down-stream signaling pathways in CRF-positive cells. Project 5 may identify CRF and PACAP systems and circuits as being fundamentally altered in stress-related illnesses such as PTSD, and is a key nexus of the Center that enhances, and is enhanced by, the other (preclinical, clinical) elements.