Upstream control and downstream effects of NPS release

说明(申请人提供)：神经肽S(NPS)是最近发现的一种脑内递质。NPS使啮齿类动物产生觉醒和觉醒，并减少焦虑的行为迹象。最近的研究还发现，NPS刺激药物寻找行为，增强学习和记忆。NPS受体基因的一种自然发生的突变与恐慌症有关，这使得NPS系统成为开发新药的有趣候选者，这些新药可能在治疗焦虑症或精神疾病方面具有治疗效果，这些疾病涉及对厌恶记忆的不当处理，如恐慌症、恐惧症或广泛性焦虑症。然而，目前尚不清楚哪些其他递质控制NPS在大脑中的释放，以及哪些递质与NPS共同定位并共同释放。为了解决这些问题，我们培育了转基因小鼠，只在那些也合成NPS的神经元中表达荧光标记蛋白(EGFP)。这些NPS/EGFP转基因小鼠使我们能够收集单个荧光神经元并确定它们的基因表达谱。在所有表达的基因中，我们对受体基因特别感兴趣，因为它们表明哪些传入的神经递质可能控制产生NPS的神经元的活动。作为这些研究的第一个结果，我们发现了神经肽CRF的功能性受体，它是调节压力和焦虑行为的关键递质。此外，我们现在有一个潜在的共同定位的其他神经递质的清单，这些神经递质可能在这些神经元被激活时共同释放。目前这项提议的第一个目标是通过用合适的抗体探针对脑切片进行染色来确认产生NPS的神经元中是否存在这些额外的神经递质。其次，我们将研究位于NPS产生细胞上的受体，通过记录NPS/EGFP转基因小鼠脑片中的电活动来激活或抑制神经元的放电。作为概念的证明，我们将重点研究CRF受体，以建立NPS系统与这个主要的应激调节系统的相互作用。最后，我们将研究应激是否导致NPS神经元的激活，特别是表达CRF受体的神经元。这种聚焦的方法将使我们能够了解NPS系统与CRF系统的功能反应和网络相互作用，并可能转化为治疗焦虑或应激相关疾病的新治疗策略。

DESCRIPTION (provided by applicant): Neuropeptide S (NPS) is a recently identified transmitter in the brain. NPS produces arousal and wakefulness and reduces behavioral signs of anxiety in rodents. Recent studies have also found that NPS stimulates drug-seeking behavior and enhances learning and memory. A naturally occurring mutation in the NPS receptor gene is associated with panic disorder, making the NPS system an interesting candidate for development of new drugs that could have therapeutic benefits in the treatment of anxiety disorders or psychiatric conditions involving improper processing of aversive memories, such as panic disorder, phobias or generalized anxiety disorder. However, it is currently unknown which other transmitters control the release of NPS in the brain and which transmitters are co-localized, and thus co-released, with NPS. To address these questions, we have generated transgenic mice that express a fluorescent marker protein (EGFP) in only those neurons that also synthesize NPS. These NPS/EGFP-transgenic mice have allowed us to collect single fluorescent neurons and determine their gene expression profile. Among all expressed genes, we are particularly interested in receptor genes as they indicate which incoming neurotransmitters may control activity of NPS-producing neurons. As a first result of these investigations, we have discovered functional receptors for the neuropeptide CRF, which is a key transmitter in mediating stress and anxiety behaviors. Furthermore, we have now a list of potentially co-localized other neurotransmitters that may be co-released whenever these neurons are activated. The first goal of the current proposal is to confirm the presence of these additional neurotransmitters in NPS-producing neurons by staining brain sections with suitable antibody probes. Secondly, we will study receptors located on NPS-producing cells that either activate or inhibit neuronal firing by recording their electrical activity in brain slices from NPS/EGFP-transgenic mice. As a proof of concept, we will focus on CRF receptors to establish the interaction of the NPS system with this major stress-regulating system. Finally, we will investigate if stress leads to activation of NPS neurons, and in particular neurons that express CRF receptors. This focused approach will allow us to understand functional responses and network interactions of the NPS system with the CRF system and could translate into novel therapeutic strategies for the treatment of anxiety or stress-related disorders.