Validation of Prenatal Rabbit Hypoxia Ischemia as a Model of Cerebral Palsy-induced Pain

项目摘要/摘要 脑性瘫痪(CP)是一种由运动障碍定义的疾病，但疼痛是最常见的并存疾病。 疼痛通常归因于肌肉僵硬、痉挛、关节痉挛、髋关节半脱位和关节炎。然而， 慢性疼痛的存在与肌肉骨骼问题的严重程度没有明确的相关性，并且可能 完全独立的(即热过敏)。CP相关性疼痛既可以是肿瘤性的，也可以是神经性的 在自然界中，它可能是由扰乱运动的相同的全局神经损伤引起的。关于疼痛的研究 在CP中，需要使用一种既表现出运动功能障碍又表现出疼痛加剧的动物模型。 我们通过验证一个更大的、非啮齿动物的CP动物模型来解决这一临床需求，该动物模型既显示了 产前缺氧后的感觉和运动障碍。新西兰白兔(Leporidae科) 具有比CP啮齿动物模型更高的表面效度、结构效度和标准效度，被广泛用于比较 慢性前列腺炎的临床前研究。我们实验室的最新数据显示，这些兔子也有增强的伤害感。我们 建议进一步验证HI兔作为慢性CP疼痛的模型，这是一个完全没有研究的领域。 目的1确定HI兔作为CP相关性疼痛模型的脸部效度。解剖学和 肌肉骨骼疼痛、神经性疼痛和神经病理性疼痛的生理学证据将在幼稚中进行评估。 对照组、HI感染组和假手术组兔从出生到性成熟(P5、P18、P60、P180)。我们 将用膜片钳技术表征伤害性感受器功能障碍的背根节神经元(10只兔的100个神经元 每个实验组的每个年龄)，外周伤害性感受器的组织学(中央传入纤维的发芽，计数 背根节内伤害性感受器的数量，以及每个实验组每个年龄6套的表皮神经纤维密度)，以及 肌肉骨骼损伤(AIMS 1-2中所有套件的改良Ashworth、关节扭矩和肌肉缩短)。 目的2验证HI兔感觉行为与CP疼痛表型的一致性。我们会 在我们关于新生儿伤害感的初步研究的基础上，验证与以下方面相关的行为进展 衰老过程中的生物-心理-社会疼痛。具体地说，我们将进行痛觉、疼痛感知、认知、 纵向测量，每个实验组每个性别20个工具包中的抑郁和焦虑样行为 与目标1相同。多变量分析将比较对照、假感染和HI影响组的措施，性别和 运动障碍的严重性。所有测试都将经过信度和效度评估。最高效、最有效的 将确定程序，以保持对疼痛检测的高灵敏度。这些测试将确定HI是否 兔子表现出健壮的、可重复的行为，表明了与患有CP的人相似的多方面疼痛。目标3 为疼痛研究确定兔慢性胰腺炎HI模型的重复性和可靠性。型号 重复性将由有经验的外科医生、护理员和行为主义者培训新手来确定。 调查人员。将在目标1-2中的所有程序和评估中评估实验者间的可靠性。 总而言之，这些测试将为HI兔在CP背景下的疼痛研究中的使用提供验证。

Project Summary / Abstract Cerebral palsy (CP) is a condition defined by motor impairment, but pain is the most prevalent co-morbidity. Pain is commonly attributed to stiff, spastic muscles, joint contractures, hip subluxation and arthritis. However, the presence of chronic pain does not neatly correlate with severity of musculoskeletal problems and may be entirely independent (ie thermal hypersensitivity). CP associated pain can be both nociplastic and neuropathic in nature and could be caused by the same global neural damage that disrupted movement. Research on pain in CP requires using an animal model that displays both phenotypes of motor dysfunction and enhanced pain. We address this clinical need by validating a larger, non-rodent animal model of CP which exhibits both sensory and motor impairments after prenatal hypoxia. New Zealand White (NZW) rabbits (family Leporidae) have higher face, construct and criterion validity than rodent models of CP and are widely used for comparative preclinical studies of CP. Recent data from our labs reveal these rabbits also have enhanced nociception. We propose to further validate HI rabbits as a model of chronic pain in CP, an area that is completely unstudied. Aim 1 Determine face validity of HI rabbits as a model of CP-associated pain. Anatomical and physiological evidence of musculoskeletal pain, nociplastic pain and neuropathic pain will be assessed in naïve control, HI affected, and sham-operated rabbit kits from birth to sexual maturity (at P5, P18, P60, P180). We will characterize nociceptor dysfunction using patch clamp of DRG neurons (n = 100 neurons from 10 rabbits per age per experimental group), histology of peripheral nociceptors (sprouting of central afferent fibers, counts of nociceptors in the DRGs, and epidermal nerve fiber density in 6 kits per age per experimental group), and musculoskeletal impairment (modified Ashworth, joint torque and muscle shortening in all kits in Aims 1-2). Aim 2 Validate that sensory behavior in HI rabbits is consistent with pain phenotypes in CP. We will build upon our initial studies on nociception in neonates to verify the progression of behaviors related to biopsychosocial pain during aging. Specifically, we will perform tests of nociception, pain perception, cognition, and depressive- and anxiety-like behaviors in 20 kits per sex per experimental group, measured longitudinally as in Aim 1. A multivariate analysis will compare measures in control, sham and HI affected groups, sex and severity of motor deficits. All tests will be assessed for reliability and validity. The most efficient and valid procedures will be identified to maintain high sensitivity for detection of pain. These tests will determine if HI rabbits show robust, reproducible behaviors indicative of the multifaceted pain similar to people with CP. Aim 3 Determine reproducibility and reliability of the rabbit HI model of CP for studies of pain. Model reproducibility will be determined by having experienced surgeons, caretakers, and behaviorists train novice investigators. Inter-experimenter reliability will be assessed in all procedures and assessments in Aims 1 - 2. Together, these tests will provide a validation of the HI rabbit for use in studies of pain in the context of CP.