Danish Headache Center, Department of Neurology, Rigshospitalet, University of Copenhagen.
Danish Headache Center, Department of Neurology, Rigshospitalet, University of Copenhagen; EHESP, Irset (Institut de Recherche en Santé, Environnement et Travail), Univ Rennes, Inserm; Department of Biology, Section of Cell Biology and Physiology, University of Copenhagen.
J Vis Exp. 2022 May 16(183). doi: 10.3791/63723.
Calcitonin gene-related peptide (CGRP) was first discovered in the 1980s as a splice variant from the calcitonin gene. Since its discovery, its role in migraine pathophysiology has been well established, first by its potent vasodilator properties and subsequently by its presence and function as a neurotransmitter in the sensory trigeminovascular system. The migraine-provoking ability of CGRP gave support to the pharma industry to develop monoclonal antibodies and antagonists inhibiting the effect of CGRP. A new treatment paradigm has proven effective in the prophylactic treatment of migraine. One of the useful tools to further understand migraine mechanisms is the ex vivo model of CGRP release from the trigeminovascular system. It is a relatively simple method that can be used with various pharmacological tools to achieve know-how to further develop new effective migraine treatments. The present protocol describes a CGRP release model and the technique to quantify the effect of pharmacological agents on the amount of CGRP released from the trigeminovascular system in rodents. A procedure describing the experimental approach from euthanasia to the measurement of protein levels is provided. The essential isolation of the trigeminal ganglion and the trigeminal nucleus caudalis from both mice and rats and the preparation of rat dura mater are described in detail. Furthermore, representative results from both species (rats and mice) are presented. The technique is a key tool to investigate the molecular mechanisms involved in migraine pathophysiology by using various pharmacological compounds and genetically modified animals.
降钙素基因相关肽（CGRP）于 20 世纪 80 年代首次被发现，是降钙素基因的剪接变体。自发现以来，其在偏头痛病理生理学中的作用已得到充分证实，首先是其强大的血管舒张特性，随后是其作为感觉三叉神经血管系统中的神经递质的存在和功能。CGRP 引起偏头痛的能力为制药行业开发抑制 CGRP 作用的单克隆抗体和拮抗剂提供了支持。一种新的治疗模式已被证明在偏头痛的预防性治疗中有效。进一步了解偏头痛机制的有用工具之一是三叉神经血管系统中 CGRP 释放的离体模型。这是一种相对简单的方法，可以与各种药理学工具一起使用，以获得专业知识，从而进一步开发新的有效偏头痛治疗方法。本方案描述了一种 CGRP 释放模型，以及一种技术，可用于定量药理学药物对从啮齿动物三叉神经血管系统释放的 CGRP 量的影响。提供了从安乐死到测量蛋白质水平的实验方法描述。详细描述了从处死到从小鼠和大鼠中分离三叉神经节和尾状核三叉神经核以及准备大鼠硬脑膜的基本程序。此外，还呈现了来自两个物种（大鼠和小鼠）的代表性结果。该技术是通过使用各种药理学化合物和基因修饰动物研究偏头痛病理生理学中涉及的分子机制的关键工具。
