School of Brain and Behavioral Sciences and Center for Advanced Pain Studies, University of Texas at Dallas.
School of Brain and Behavioral Sciences and Center for Advanced Pain Studies, University of Texas at Dallas;
J Vis Exp. 2021 Jul 29(173). doi: 10.3791/62867.
The cranial meninges, comprised of the dura mater, arachnoid, and pia mater, are thought to primarily serve structural functions for the nervous system. For example, they protect the brain from the skull and anchor/organize the vascular and neuronal supply of the cortex. However, the meninges are also implicated in nervous system disorders such as migraine, where the pain experienced during a migraine is attributed to local sterile inflammation and subsequent activation of local nociceptive afferents. Of the layers in the meninges, the dura mater is of particular interest in the pathophysiology of migraines. It is highly vascularized, harbors local nociceptive neurons, and is home to a diverse array of resident cells such as immune cells. Subtle changes in the local meningeal microenvironment may lead to activation and sensitization of dural perivascular nociceptors, thus leading to migraine pain. Studies have sought to address how dural afferents become activated/sensitized by using either in vivo electrophysiology, imaging techniques, or behavioral models, but these commonly require very invasive surgeries. This protocol presents a method for comparatively non-invasive application of compounds on the dura mater in mice and a suitable method for measuring headache-like tactile sensitivity using periorbital von Frey testing following dural stimulation. This method maintains the integrity of the dura and skull and reduces confounding effects from invasive techniques by injecting substances through a 0.65 mm modified cannula at the junction of unfused sagittal and lambdoid sutures. This preclinical model will allow researchers to investigate a wide range of dural stimuli and their role in the pathological progression of migraine, such as nociceptor activation, immune cell activation, vascular changes, and pain behaviors, all while maintaining injury-free conditions to the skull and meninges.
颅脑膜由硬脑膜、蛛网膜和软脑膜组成，被认为主要为神经系统提供结构功能。例如，它们保护大脑免受颅骨的伤害，并固定/组织大脑皮层的血管和神经元供应。然而，脑膜也与偏头痛等神经系统疾病有关，在偏头痛期间经历的疼痛归因于局部无菌性炎症和随后的局部伤害性传入纤维的激活。在脑膜的各层中，硬脑膜在偏头痛的病理生理学中特别重要。它高度血管化，含有局部伤害性神经元，并且是各种常驻细胞（如免疫细胞）的家园。局部脑膜微环境的细微变化可能导致脑膜血管周围伤害感受器的激活和敏化，从而导致偏头痛疼痛。研究人员试图通过使用体内电生理学、成像技术或行为模型来解决硬脑膜传入神经如何被激活/敏化的问题，但这些方法通常需要非常侵入性的手术。本方案提出了一种在小鼠硬脑膜上比较非侵入性应用化合物的方法，以及一种在硬脑膜刺激后使用眶周 von Frey 测试测量类似头痛的触觉敏感性的合适方法。该方法通过在未融合的矢状缝和人字缝交界处的 0.65 毫米改良套管内注射物质，保持硬脑膜和颅骨的完整性，并减少侵入性技术的混杂影响。这种临床前模型将使研究人员能够研究广泛的硬脑膜刺激及其在偏头痛病理进展中的作用，如伤害感受器激活、免疫细胞激活、血管变化和疼痛行为，同时保持颅骨和脑膜不受损伤。
