Targeted regulation of CGRP activity in trigeminovascula

这些研究的主要目的是研究降钙素基因4“相关肽(CGRP)在三叉神经血管系统中活性的分子控制。三叉神经节神经元的降钙素基因相关肽是脑循环中最有效的血管扩张机制之一。降钙素基因相关肽在蛛网膜下腔出血、高血压和偏头痛中的作用突出。我们最近发现，病毒注射降钙素基因相关肽能够预防蛛网膜下腔出血后的血管痉挛。为了了解降钙素基因相关肽的作用机制，我们现在建议研究降钙素基因相关肽受体调节亚基。三叉神经节神经元和脑血管上既有突触前受体也有突触后受体的报道。然而，定义CGRP受体活性的分子机制直到最近才为人所知。CGRP受体是降钙素受体样受体(CRLR)的三聚体和两个调节亚基，分别称为受体活性修饰蛋白1(RAMP1)和受体组件蛋白(RCP)。 RAMP1是配体结合活动所必需的，而RCP是完整信号活动所必需的。我们建议验证CGRP受体亚单位的基因转移可以增加三叉神经血管系统中CGRP活性的假设。具体地说，我们将询问RAMP1和RCP蛋白是否可以调节三叉神经节神经元和血管肌肉中突触前和突触后CGRP受体的功能。方法将是使用原代细胞培养和转基因小鼠。首先，将在培养的三叉神经细胞和血管中测试CGRP受体蛋白 肌肉。其次，将使用RAMP1和RCP转基因小鼠的大动脉在体外测量CGRP诱导的血管松弛，并将在体内研究脑微血管的血管运动反应。第三，在野生型和转基因小鼠中，结合CGRP基因转移，将比较基底动脉的松弛。该项目建立并继续了我们过去关于CGRP和CGRP基因转移在脑血管系统中的作用的研究。通过 将受体研究与我们的编码CGRP的腺病毒载体相结合，我们期望为CGRP基因传递和CGRP受体调控的联合方法奠定基础。该项目将汇集脑循环(Faraci和Heistad)、CGRP表达(RUSSO)和CGRP受体功能(Dickerson)方面的专业知识。该项目的意义在于它将提供对CGRP作用机制的新的理解和潜在的新的 对血管疾病治疗策略的展望。

The major goal of these studies is to examine the molecular control of calcitonin gene4"elated peptide (CGRP) activity in the trigeminovascular system. CGRP from trigeminal ganglia neurons is one of the most potent vasodilator mechanisms in the cercbral circulation. The importance of CGRP is highlighted by its involvement in subarachnoid hemorrhage, hypertension, and migraine. We have recently shown that viral delivery of CGRP is capable of preventing vasospasm after subarachnoid hemorrhage. To understand the mechanisms of CGRP action, we now propose to study the CGRP receptor regulatory subunits. Both presynaptic and postsynaptic receptors on the trigeminal ganglia neurons and cerebral vessels have been reported. However, the molecular mechanisms that define CGRP receptor activity are only recently becoming known. The CGRP receptor is a trimer of the calcitonin receptor-like receptor (CRLR) and two regulatory subunits called receptor activity-modifying protein 1 (RAMP1) and receptor component protein (RCP). RAMP1 is required for ligand binding activity and RCP is required for full signaling activity. We propose to test the hypothesis that gene transfer of CGRP receptor subunits can increase CGRP activity in the trigeminovascular system. Specifically, we will ask whether the RAMP1 and RCP proteins can regulate presynaptic and postsynaptic CGRP receptor function in trigeminal ganglia neurons and vascular muscle. The approach will be to use primary cell cultures and transgenic mice. First, the CGRP receptor proteins will be tested in cultured trigeminal neurons and vascular muscle. Second, CGRP-induced vasorelaxation will be measured in vitro using large arteries from RAMP1 and RCP transgenic mice and vasomotor responses will be studied in cerebral microvessels in vivo. Third, relaxation of the basilar artery will be compared in wildtype and transgenic mice in combination with CGRP gene transfer. This project builds on and continues our past studies on effects of CGRP and CGRP gene transfer in the cerebrovasculature. By coupling receptor studies with our adenoviral vector encoding CGRP we expect to lay the foundation for a combined approach of CGRP gene delivery and CGRP receptor modulation. The project will bring together expertise in cerebral circulation (Faraci and Heistad), CGRP expression (Russo), and CGRP receptor function (Dickerson). The significance of this project is that it will provide a new understanding of mechanisms of CGRP action and a potentially new perspective on therapeutic strategies for vascular disorders.