Expression of the CGRP Family of Neuropeptides and their Receptors in the Trigeminal Ganglion
Expression of the CGRP Family of Neuropeptides and their Receptors in the Trigeminal Ganglion
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DOI：
10.1007/s12031-020-01493-z
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发表时间：
2020-02-22
影响因子：
3.1
通讯作者：
Warfvinge, Karin
中科院分区：
文献类型：
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作者：
Edvinsson, Lars;Grell, Anne-Sofie;Warfvinge, Karin
降钙素基因相关肽（CGRP）家族的神经肽，由CGRP，肾上腺髓质素，胰淀素，和降钙素。这些受体由降钙素受体样受体（CLR）或降钙素受体（CTR）组成，其功能需要辅助蛋白质，即受体活性修饰蛋白（RAMP）。CGRP在原发性头痛中具有关键作用，但CGRP肽家族的其他成员在头痛中的作用尚不清楚。在这里，我们描述了这些分子在三叉神经节（TG）中的表达，以了解更多关于它们可能的作用。使用抗CGRP、降钙素原、降钙素、肾上腺髓质素、胰淀素、RAMP 1/2/3、RAMP和CTR的一抗，对大鼠TG的冷冻切片进行单或双免疫组化。此外，通过TG上的定量qPCR测量mRNA表达。CGRP和降钙素在中小神经元胞质中表达丰富，有时共定位。在胶质细胞中观察到降钙素原。还观察到储存CGRP和降钙素的免疫反应性纤维。肾上腺髓质素免疫反应被发现在卫星胶质细胞和纤维，可能的髓鞘雪旺细胞。胰淀素在许多TG神经元的细胞质中被发现。使用qPCR测量肾上腺髓质素、胰淀素、RAMP、RAMP 1、RAMP 2、RAMP 3和CTR的mRNA表达水平。实验验证了TG中mRNA的表达，CTR除外，其高于检测限，表明很少或没有mRNA表达。除了已知的CGRP受体（CGRP/RAMP 1）和降钙素-CTR受体外，我们认为大鼠TG中还存在其他受体：肾上腺髓质素受体AM（2）（CGRP/RAMP 3）主要存在于卫星胶质细胞中，胰淀素受体AMY（1）（CTR/RAMP 1）主要存在于神经元中，AMY（3）（CTR/RAMP 3）主要存在于卫星胶质细胞中。重要的是在研究中并排比较肽和受体，以帮助解决由受体之间的交叉反应性引起的作用问题。CGRP肽家族的多种生物学作用中的几种是临床相关的。我们的研究结果表明，特定的配体和受体网站在大鼠三叉神经节，突出识别机制，以促进药物开发。
The calcitonin gene-related peptide (CGRP) family of neuropeptides, consists of CGRP, adrenomedullin, amylin, and calcitonin. The receptors consist of either calcitonin receptor-like receptor (CLR) or calcitonin receptor (CTR) which for function needs an accessory protein, receptor activity-modifying proteins (RAMPs). CGRP has a pivotal role in primary headaches but the role of the other members of the CGRP family of peptides in headaches is not known. Here, we describe the expression of these molecules in the trigeminal ganglion (TG) to understand more on their possible role(s). Single or double immunohistochemistry were applied on frozen sections of rat TG using primary antibodies against CGRP, procalcitonin, calcitonin, adrenomedullin, amylin, RAMP1/2/3, CLR, and CTR. In addition, mRNA expression was measured by quantitative qPCR on TGs. CGRP and calcitonin showed rich expression in the cytoplasm of small to medium-sized neurons, and co-localized sometimes. Procalcitonin was observed in the glial cells. Immunoreactive fibers storing both CGRP and calcitonin were also observed. Adrenomedullin immunoreactivity was found in the satellite glial cells and in fibers, probably the myelinating Schwann cells. Amylin was found in the cytoplasm in many TG neurons. Levels of mRNA expression for adrenomedullin, amylin, CLR, RAMP1, RAMP2, RAMP3, and CTR were measured using qPCR. The experiments verified the expression of mRNA in the TG with the exception of CTR, which was above the limit of detection indicating little or no mRNA expression. In addition to the well-known CGRP receptor (CLR/RAMP1) and the receptor for calcitonin-CTR, we propose that other receptors exist in the rat TG: adrenomedullin receptor AM(2) (CLR/RAMP3) in mainly the satellite glial cells, amylin receptors AMY(1) (CTR/RAMP1) in mainly neurons, and AMY(3) (CTR/RAMP3) in the satellite glial cells. It is important to compare peptides and receptors side-by-side in studies to help address questions of actions resulting from cross-reactivity between receptors. Several of the diverse biological actions of the CGRP family of peptides are clinically relevant. Our findings demonstrate the specific ligand and receptor sites in the rat trigeminal ganglion, highlighting recognition mechanisms to facilitate drug development.