Spatiotemporal Coding in the Pain Circuit Along the Spine-brain Continuum

沿着脊柱-大脑连续体的疼痛回路的时空编码

• 批准号: 10205394
• 负责人: David Allenson Borton
• 金额: $ 8.96万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10205394
• 关键词: Acute Pain; Afferent Neurons; Animals; BRAIN initiative; Behavior; Behavioral; Brain; Calcium; Cations; Cells; Cellular biology; Chronic; Code; Data; Detection; Electrophysiology (science); Experimental Designs; Family; Fiber; Future; Goals; Grant; Head; Health Sciences; Image; Implant; In Vitro; Interneurons; Intervention; Investigation; Light; Medical; Medicine; Methodology; Methods; Microelectrodes; Microscope; Microscopy; Modeling; Monitor; Mus; Nerve Fibers; Neuraxis; Neurons; Neurosciences; Nociception; Optics; Outcome; Pain; Pain intensity; Parents; Parvalbumins; Perception; Peripheral; Physicians; Posterior Horn Cells; Process; Reflex action; Reporting; Research; Rest; Scientist; Sensory; Signal Transduction; Site; Spinal; Spinal Cord; Spinal cord posterior horn; TRPV1 gene; Tactile; Techniques; Thalamic structure; Time; Touch sensation; Training; Transgenic Mice; Underrepresented Populations; Universities; Vertebral column; Video Recording; Viral; Work; afferent nerve; awake; base; calmodulin-dependent protein kinase II; career; cell type; chronic pain; detection assay; dorsal horn; excitatory neuron; experience; extracellular; improved; in vivo; medical schools; member; millisecond; neurophysiology; optogenetics; pain processing; pain signal; receptor; relating to nervous system; response; skills; spatiotemporal; transmission process

PROJECT SUMMARY Understanding the cellular biology and neurophysiology of sensory processing in the spinal cord is fundamental to advancing medical intervention in the treatment of chronic and acute pain conditions. The current understanding of the neurophysiology of spinal cord circuitry is founded on experimental single-unit electrophysiology on anesthetized animals and in-vitro studies, but limited data exist from in-vivo functional circuitry of sensory signals. Part of the challenge to such experimentation has been the limited capacity for monitoring electrophysiologic signals in awake animals and inducing reliable activation of pain fibers. Consequently, the activity of specific neuronal subtypes in propagating excitatory and inhibitory signals involved in the transmission of pain signals remains unknown in-vivo. Recently, we have developed a pain detection assay consisting of a lick behavior in response to optogenetic activation of predominantly nociceptive peripheral afferent nerve fibers in head-restrained transgenic mice expressing Channelrhodopsin 2 (ChR2) in transient receptor potential cation channel subfamily V member 1 (TRPV1) containing neurons. In this model, mice are trained to provide lick reports to the detection of light-evoked nociceptive stimulation to the hind paw. Our nociceptive lick-report detection assay enables a host of investigations into the millisecond, single-cell, neural dynamics underlying pain processing in the central nervous system of awake behaving animals. Further, we have developed a “backpack drive” to provide multi-site chronic extracellular recordings from dorsal horn neurons derived from superficial laminas II-III. Unfortunately, such electrophysiology cannot be used to determine cellular subclasses during recording. Here, we will focus on advancing our ability to record cell-type-specific activity in the dorsal horn in response to light-activated TRPV1 containing neurons in the periphery. We will develop a reliable method for achieving consistent GCaMP6-family expression in specific neuronal cell types (e.g. CaMKII, PV) involved in the specific activation of pain signals through our optogenetic stimulation experimental design. We will optimize a spinal optical window to perform awake Calcium imaging during time-locked tactile input and characterize calcium dynamics in neuronal subtypes in the dorsal horn during behavioral tasks. This work will establish a methodology to collect temporal dynamics of large classes of neurons in the dorsal horn in response to time-locked, spatially-precise, and amplitude-modulated input in the periphery leading to improved understanding of acute pain conditions.

项目总结 了解脊髓感觉处理的细胞生物学和神经生理学 这是推进治疗慢性和急性疼痛的医疗干预的基础。这个 目前对脊髓环路神经生理学的认识是建立在实验单一单位的基础上的 麻醉动物的电生理学和体外研究，但来自体内功能的数据有限 感官信号的回路。这种实验面临的部分挑战是有限的能力 监测清醒动物的电生理信号，并诱导疼痛纤维的可靠激活。 因此，特定神经元亚型在传播兴奋性和抑制性信号时的活动 在体内，痛觉信号的传递机制尚不清楚。最近，我们出现了一种疼痛 由对主要伤害性感受器的光基因激活作出反应的舔行为组成的检测方法 表达通道视紫红质2基因小鼠的外周传入神经纤维 含有神经元的瞬时受体电位阳离子通道亚家族V成员1(TRPV1)。在这个模型中， 小鼠被训练来提供舔舔报告，以检测到光对后爪的伤害性刺激。 我们的伤害性舔报告检测方法能够对毫秒级、单细胞、 清醒行为动物中枢神经系统疼痛处理的神经动力学。 此外，我们还开发了一种“背包驱动器”来提供多个地点的慢性细胞外录音。 来自浅层板层的背角神经元II-III。不幸的是，这种电生理学不能 用于在记录过程中确定蜂窝亚类。在这里，我们将重点提高我们的记录能力 背角细胞类型特异性活动对光激活的含TRPV1神经元的反应 外围设备。我们将开发一种可靠的方法来实现GCaMP6家族在特定情况下的一致表达 神经细胞类型(如CaMKII、PV)通过我们的光发生参与痛觉信号的特定激活 刺激性实验设计。我们将优化一个脊髓光学窗口来进行清醒的钙成像 时间锁定触觉输入过程中背角神经元亚型的钙动力学特征 在行为任务中。这项工作将建立一种方法来收集大班的时间动力学 背角神经元对时间锁定、空间精确和幅度调制的输入的反应 外周刺激有助于改善对急性疼痛状况的了解。