Understanding spinal neuropeptide signaling in itch

项目摘要/摘要 慢性瘙痒疾病是去看皮肤科医生的主要原因，对患者有毁灭性的影响 生活质量。尽管进行了广泛的研究努力，但在一定程度上，治疗慢性瘙痒的有效方法仍然难以找到。 因为人们对瘙痒的基本神经编码知之甚少。调查员的长期目标是 研究计划是阐明在正常和正常情况下处理瘙痒输入的神经机制 病理情况。有趣的是，慢性瘙痒障碍的一个特征是兴奋性的上调。 在脊髓内驱动瘙痒的神经肽-P物质(SP)和胃泌素释放肽(GRP)。 然而，神经肽调节脊髓回路产生瘙痒的机制尚不清楚。致信地址 这一认识上的根本差距，这一建议将调查神经肽(SP和GRP)是否在 与神经递质平行(在相同的靶点上)，或者神经肽信号与 神经传递(参与不同的靶点)，从而重新配置脊髓回路。具体地说，调查员 将检验这一假说，即引起瘙痒的神经肽SP和GRP通过不同的信号作用于 重新配置急性和持续性瘙痒的神经回路。在K99阶段，她将学习和应用切割- 从神经元超微结构到群体水平的边缘方法。目标1(K99)将比较脊柱 用多重电子显微镜观察突触与神经肽的连接性。目标2(K99)将 识别神经肽激活的脊髓网络与SP(和GRP)脊髓的突触传递 使用双光子钙成像结合体外体感准备的神经元。一部分 K99阶段还将致力于学习实现机器学习算法来检测和量化 老鼠瘙痒行为，以及职业发展活动，如获得领导力和指导技能以及 在会议上发表演讲，以扩大申请人的职业关系网。目标3(R00)将决定是否 脊髓异常自发活动(AIM)的表现需要SP和GRP信号 3A)和与使用药物抑制的持续性瘙痒相关的提高瘙痒行为(目标3B 战略。这些研究将利用她在双光子钙离子成像和自动行为方面的新专业知识 结合她在啮齿动物瘙痒方面的强大背景，分析在K99阶段开发的管道 模特们。总之，这些实验将对脊髓瘙痒中的神经肽信号提供基本的见解。 传递和确定脊髓神经肽信号在持续性瘙痒中的必要性。这件事的成功 职业奖励培训计划，包括科学和职业发展活动，将由 匹兹堡疼痛中心提供的科学专业知识、协作和教育机会 研究。完成这项计划将使申请者为领导创新研究项目做好准备 独立调查员。

PROJECT SUMMARY/ABSTRACT Chronic itch disorders are the primary reason for visits to dermatologists and have devasting effects on patient quality of life. Despite extensive research efforts, an effective treatment for chronic itch remains elusive, in part because the basic neural coding that underlies itch is poorly understood. The long-term goal of the investigator’s research program is to elucidate the neural mechanisms that underlie processing of itch input under normal and pathological conditions. Interestingly, one hallmark of chronic itch disorders is the upregulation of excitatory neuropeptides that drive itch – substance P (SP) and gastrin-releasing peptide (GRP) – within the spinal cord. However, the mechanism by which neuropeptides modulate spinal circuits to produce itch is unclear. To address this fundamental gap in knowledge, this proposal will investigate whether neuropeptides (SP and GRP) act in parallel with neurotransmitters (on the same targets) or if neuropeptide signaling diverges from neurotransmission (engage distinct targets), thereby reconfiguring spinal circuits. Specifically, the investigator will test the hypothesis that the itch-inducing neuropeptides SP and GRP act via divergent signaling to reconfigure neural circuits in acute and persistent itch. During the K99 Phase, she will learn and apply cutting- edge approaches that span from the neuron ultrastructural to population level. Aim 1 (K99) will compare spinal synaptic connectivity to neuropeptide connectivity using multiplexed electron microscopy. Aim 2 (K99) will identify the spinal networks activated by neuropeptide versus synaptic transmission from SP (and GRP) spinal neurons using two-photon Ca2+ imaging in combination with an ex vivo somatosensory preparation. A portion of the K99 phase will also be dedicated to learning to implement machine learning algorithms to detect and quantify mouse itch behaviors, and to career development activities such as gaining leadership and mentorship skills and presenting at conferences to expand the applicant’s professional network. Aim 3 (R00) will determine whether the SP and GRP signaling are required for the manifestation of aberrant spontaneous spinal cord activity (Aim 3A) and elevated itch behaviors (Aim 3B) associated with persistent itch using pharmacological inhibition strategies. These studies will leverage her new expertise in two-photon Ca2+ imaging and automated behavioral analysis pipelines developed during the K99 phase in combination with her strong background in rodent itch models. Together, these experiments will provide fundamental insights into neuropeptide signaling in spinal itch transmission and identify the necessity of spinal neuropeptide signaling in persistent itch. The success of this career award training plan, which includes activities for scientific and career development, will be aided by the scientific expertise, collaboration, and educational opportunities offered by the Pittsburgh Center for Pain Research. Completing this plan will prepare the applicant to lead an innovative research program as an independent investigator.