Developmental Regulation of Intrinsic Excitability in Spinal Pain Networks

描述（由申请人提供）：尽管很明显，即使是最小的婴儿也会因损伤、疾病、手术或重症监护治疗而经历明显的疼痛，但对于组织损伤如何影响中枢神经系统（CNS）疼痛回路中神经元的内在兴奋性，我们知之甚少。由于脊髓浅背角（SDH）在疼痛通路中起着关键的中继站作用，因此更好地了解不同年龄的组织损伤如何影响SDH神经元的内在膜特性是解决这一问题的重要且合乎逻辑的第一步。长期目标是通过确定更适合发育的新型镇痛策略来改善婴儿和儿童疼痛的临床治疗。该应用程序的总体目标是确定在正常和病理条件下调节SDH内发育神经元内在兴奋性的关键离子电导，这是追求该目标的下一步。核心假设是，在出生后早期发育的关键时期，I层内的兴奋性中间神经元表现出内在的起搏器型振荡，这是由持续的Na+和Ca2+电流驱动的，并由新生儿组织损伤促进。这项研究的基本原理是，理解内在神经元兴奋性是如何在未成熟的伤害性回路中被特异性调节的，将揭示调节其输出的新方法，这在成人的研究中是不明显的。在强大的初步数据的指导下，将对中心假设进行测试，并通过追求以下具体目标来实现本应用程序的总体目标：1)确定在出生后早期发育过程中自发活跃的SDH神经元；(2)阐明驱动新生儿SDH神经元内禀放电的离子机制；(3)检测早期组织损伤后SDH网络中离子通道表达和固有兴奋性的细胞类型特异性变化。这些目标将通过使用体外电生理、免疫组织化学和生化技术来表征发育中的SDH神经元的确定亚型中调节神经元兴奋性的离子机制，并确定这些内在膜特性在多大程度上受组织损伤以年龄依赖的方式调节。这些调查的结果将是对未成熟的脊柱疼痛网络如何在细胞水平上控制活动的新见解。因此，提出的研究是重要的，因为它将开始提供所需的知识，以发展证据为基础的治疗慢性儿童疼痛。

DESCRIPTION (provided by applicant): Although it is clear that even the youngest infants can experience significant pain as the result of injury, disease, surgery or intensive care therapy, little is known about how tissue damage affects the intrinsic excitability of neurons within developing pain circuits in the central nervous system (CNS). Since the superficial dorsal horn of the spinal cord (SDH) functions as a critical relay station in the pain pathway, a better understanding of how the intrinsic membrane properties of SDH neurons are influenced by tissue injury at different ages represents an important and logical first step towards addressing this issue. The long-term goal is to improve the clinical treatment of pain in infants and children by identifying novel analgesic strategies which are more developmentally appropriate. The overall objective of this application, which is the next step in pursuit of that goal, is to identify the key ionic conductances which regulate the intrinsic excitability of developing neurons within the SDH under normal and pathological conditions. The central hypothesis is that excitatory interneurons within lamina I exhibit intrinsic, pacemaker-type oscillations during a critical period of early postnatal development, which are driven by persistent Na+ and Ca2+ currents and facilitated by neonatal tissue damage. The rationale of the proposed research is that understanding how intrinsic neuronal excitability is specifically regulated within immature nociceptive circuits will reveal new ways to modulate their output which would not be evident from studies in the adult. Guided by strong preliminary data, the central hypothesis will be tested and the overall objective of this application achieved by pursuing the following specific aims: 1) Identify the SDH neurons which are spontaneously active during early postnatal development; (2) Elucidate the ionic mechanisms which drive the intrinsic firing of neonatal SDH neurons; and (3) Detect cell-type-specific changes in ion channel expression and intrinsic excitability within the developing SDH network following early tissue damage. These aims will be accomplished by using in vitro electrophysiological, immunohistochemical and biochemical techniques to characterize the ionic mechanisms regulating neuronal excitability in identified subtypes of developing SDH neurons and determine the extent to which these intrinsic membrane properties are modulated by tissue injury in an age-dependent manner. The outcome of these investigations will be new insight into how activity within immature spinal pain networks is controlled at the cellular level. As a result, the proposed research is significant because it will begin to provide the knowledge needed to develop evidence- based treatments for chronic pediatric pain. PUBLIC HEALTH RELEVANCE: The expected outcomes of the proposed research will have a positive impact on public health by illustrating that the underlying causes of pain hypersensitivity under pathological conditions are highly developmentally regulated. By elucidating the mechanisms by which early tissue damage modulates intrinsic neuronal excitability within central nociceptive networks during postnatal development, this research also brings the promise of identifying novel approaches to regulate pain pathways in an age-specific manner, which would greatly aid efforts to improve the clinical management of pediatric pain.