The neurovascular structures in the cranial dura mater have been studied with various histological techniques in the past years. In order to obtain a proper approach to reveal the detailed structures, different labeling methods for the cranial vessels and nerve fibers were tested in this study. Firstly, the labeling characteristics of phalloidin, alpha smooth muscle actin (α-SMA), and CD31 were compared in rat whole-mount cranial dura mater by using fluorescent immunohistochemistry or histochemistry. Secondly, according to their properties, phalloidin and α-SMA were selected to combine with calcitonin gene-related peptide (CGRP) to further demonstrate the cranial neurovascular structure. By these approaches, a three-dimensional map of blood vessels and nerve fibers within the whole-mount rat cranial dura mater was obtained. The results showed that phalloidin, α-SMA, and CD31 were preferably expressed in the wall of cranial vessels, corresponding to the arteriors, venules, and capillaries, respectively. Additionally, CGRP + nerve fibers were clearly demonstrated together with phalloidin + or α-SMA + vessels, forming a delicate neurovascular network in the cranial dura mater. The thick nerve bundles ran closely to the phalloidin + or α-SMA + vessels in parallel pattern, while the thin nerve fibers branched off from the bundles tending to surround the phalloidin + arterioles rather than α-SMA + venules. These findings suggest that phalloidin could be an appropriate biochemical maker to be effectively used together with CGRP for experiments examining the detailed spatial correlation of cranial blood vessels and nerve fibers in a three-dimensional view, which may provide clues for understanding the underlying mechanisms of cranial neurovascular disorders.

近年来，已通过各种组织学技术研究了颅硬脑膜的神经血管结构。为了获得适当的方法来揭示详细的结构，在这项研究中测试了颅脑血管和神经纤维的不同标记方法。首先，通过荧光免疫组织化学或组织化学方法比较了鬼笔环肽，α平滑肌肌动蛋白（α-SMA）和CD31在大鼠全颅硬脑膜中的标记特征。其次，根据其性质，选择鬼笔环肽和α-SMA与降钙素基因相关肽（CGRP）结合，进一步证明颅神经血管结构。通过这些方法，获得了整个安装的大鼠颅硬脑膜内血管和神经纤维的三维图。结果表明，鬼笔环肽，α-SMA和CD31优选在颅血管壁中表达，分别对应于前房，小静脉和毛细血管。此外，CGRP +神经纤维与鬼笔环肽+或α-SMA+血管清楚地结合在一起，在颅硬脑膜中形成了精致的神经血管网络。粗大的神经束以平行的方式紧密贴着鬼笔环肽+或α-SMA+血管，而细神经纤维从束中分叉，倾向于围绕鬼笔环肽+小动脉而不是α-SMA+小静脉。这些发现表明，鬼笔环肽可能是一种合适的生化试剂，可以与CGRP一起有效地用于实验，以三维视角检查颅脑血管和神经纤维的详细空间相关性，