TRIGEMINAL NERVE--CONTROL OF THE BRAIN VASCULATURE

建议进行实验，以建立更合理的药物基础 偏头痛和相关头痛的发现。 这个调查领域 在过去的11年里一直得到国家卫生研究院的支持。 偏头痛，第二次 失业的主要原因，困扰着高达15%的人， 女性占优势（3：1）。 抗偏头痛药物（例如，舒马曲坦和 丙戊酸盐）减少啮齿动物硬脑膜内的神经源性炎症（NI）， 降低三叉神经尾侧核（TNC）内c-fos免疫反应性 是因为有害的脑膜刺激 舒马曲坦及相关化合物 可能通过作用于5-HT 1D受体来减轻头痛， 三叉神经纤维或血管平滑肌（或两者）上。 目的1中的实验旨在定义存在/不存在， 三叉神经纤维上5-HT 1B/D受体亚型对NI和c- fos表达。 在没有区分5- HT1Dalpha\beta，我们建议使用缺乏5-HT 1B或5-HT 1D的敲除小鼠。 HT 1D α受体，以解决NI，c-fos中这些位点的相关性 表达和软脑膜血管反应使用颅窗。 我们也 建议使用反义策略来测试5-HT 1D α mRNA是否 阻断抑制激动剂对NI或c-fos表达的作用， 有害的脑膜刺激 我们建议扩大 通过比较受体结合和基因表达， 在啮齿动物和人类三叉神经节和相关死后组织中 应用受体放射自显影、原位杂交和RT-PCR 技术. 具体目标二将审查最近提出的 偏头痛治疗中的治疗靶点（GABAA）并扩展初步数据 表明丙戊酸盐（阻断GABA降解）阻断了 通过荷包牡丹碱敏感的机制。 研究将确定 受体是否在血脑屏障（BBB）外（如5- HT 1D），并通过这样做，提出了替代治疗方法， BBB不渗透性GABA能药物。 我们将扩大初步数据显示， GABAA激动剂和调节剂（即，神经类固醇，和 苯二氮卓类）依赖于副交感神经的完整性 支配脑膜 分子策略，包括原位 将使用杂交和RT-PCR来确认亚型并建立 GABA受体在人类和啮齿动物脑膜中的位置。

Experiments are proposed to establish a more rational basis for drug discovery in migraine and related headaches. This area of investigation has been supported by the NIH for the past 11 years. Migraine, the 2nd leading cause of work loss, afflicts up to 15% of people with a predominance of females (3:1). Antimigraine drugs (e.g., sumatriptan and valproate) reduce neurogenic inflammation (NI) within rodent dura mater and decrease c-fos immunoreactivity within trigeminal nucleus caudalis (TNC) following noxious meningeal stimulation. Sumatriptan and related compounds might decrease headache by acting on 5-HT1D receptors located either on trigeminal nerve fibers or on vascular smooth muscle (or both). Experiments in Aim 1 are intended to define the presence/absence and importance of 5-HT1B/D receptors subtypes on trigeminal fibers to NI and c- fos expression. In the absence of drugs which discriminate between 5- HT1Dalpha\beta, we propose to use knockout mice lacking 5-HT1B or 5- HT1Dalpha receptors to address the relevance of these sites in NI, c-fos expression, and pial vasomotor responses using a cranial window. We also propose to use antisense strategies to test whether 5-HT1Dalpha mRNA blockade inhibits the effects of agonists on NI or c-fos expression after noxious meningeal stimulation. We propose to extend the relevance of rodent data to humans by comparing receptor binding and gene expression within rodent and human trigeminal ganglions and related postmortem tissues using receptor autoradiography, in situ hybridization, and RT-PCR techniques. Specific Aim II will examine the possible importance of a recently proposed therapeutic target in migraine therapy (GABAA) and extend preliminary data showing that valproate (which blocks GABA degradation) blocks NI within the meninges by bicuculline-sensitive mechanisms. Studies will determine whether the receptor is outside the blood brain barrier (BBB) (like 5- HT1D), and by so doing, suggest alternative therapeutic approaches using BBB impermeant GABAergic drugs. We will extend preliminary data showing that GABAA agonists and modulators (i.e., neurosteroids, and benzodiazepines) are dependent upon the integrity of parasympathetic nerves innervating the meninges. Molecular strategies including in situ hybridization and RT-PCR will be used to confirm the subtype and establish the location of GABA receptors within human and rodent meninges.