Upstream control and downstream effects of NPS release

描述（由申请人提供）：神经肽S（Neuropeptide S）是最近在大脑中发现的一种递质。在啮齿类动物中，抗抑郁药产生唤醒和觉醒，并减少焦虑的行为迹象。最近的研究还发现，多巴胺能刺激觅药行为，增强学习和记忆。一个自然发生的突变，在神经元受体基因与恐慌症，使神经元系统的一个有趣的候选人，为开发新的药物，可能有治疗效益，在治疗焦虑症或精神疾病涉及不当处理的厌恶性记忆，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大脑中的β-内酰胺酶的释放，以及哪些递质与β-内酰胺酶共定位，从而共同释放。为了解决这些问题，我们已经产生了转基因小鼠，只在那些也合成荧光标记蛋白的神经元中表达荧光标记蛋白（EGFP）。这些GFP/EGFP转基因小鼠使我们能够收集单个荧光神经元并确定其基因表达谱。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的神经元的活性。作为这些研究的第一个结果，我们发现了神经肽CRF的功能受体，CRF是介导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列表，这些神经元被激活时可能会共同释放。目前建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这些额外的神经递质在产生NPS的神经元中的存在。其次，我们将研究位于NPS产生细胞上的受体，这些受体通过记录它们在转基因小鼠脑切片中的电活动来激活或抑制神经元放电。作为一个概念的证明，我们将集中在CRF受体，以建立与这一主要的压力调节系统的相互作用的CRF系统。最后，我们将研究压力是否会导致神经元的激活，特别是表达CRF受体的神经元。这种集中的方法将使我们能够了解功能性反应和网络的相互作用，CRF系统的神经系统，并可以转化为新的治疗策略，用于治疗焦虑或压力相关的疾病。 公共卫生相关性：神经元系统是治疗焦虑症和涉及厌恶性记忆异常处理的精神疾病的新兴靶点，如惊恐障碍、恐惧症或广泛性焦虑症。目前尚不清楚哪些递质控制内源性β-内酰胺酶的释放，哪些递质与β-内酰胺酶共同释放。初步的数据表明CRF介导的激活的神经元，从而连接的神经元系统与大脑的主要应激调节系统。目前的建议将详细研究这种相互作用，以设计新的治疗策略来治疗焦虑或应激障碍。

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. PUBLIC HEALTH RELEVANCE: The NPS system is an emerging target for the treatment of anxiety disorders and psychiatric conditions involving aberrant processing of aversive memory, such as panic disorder, phobias, or generalized anxiety disorder. It is currently unknown which transmitters control release of endogenous NPS and which transmitters are co-released together with NPS. Preliminary data suggest CRF-mediated activation of NPS neurons, thus linking the NPS system with the brain's major stress-modulatory system. The current proposal will study this interaction in detail in order to design novel therapeutic strategies to treat anxity or stress disorders.