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A new approach for examining the neurovascular structure with phalloidin and calcitonin gene-related peptide in the rat cranial dura mater

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Abstract

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.

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Funding

This study was supported by the project of National Key R&D Program of China (No. 2019YFC1709103; No. 2018YFC1707804), National Natural Science Foundation of China (No. 81774211; 81774432; 81801561).

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Authors and Affiliations

  1. Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, Beijing, 100700, China

    Jia Wang, Dongsheng Xu, Jingjing Cui, Shuya Wang, Chen She, Hui Wang, Shuang Wu, Jianliang Zhang, Bing Zhu & Wanzhu Bai

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  1. Jia Wang
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  2. Dongsheng Xu
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  3. Jingjing Cui
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  4. Shuya Wang
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  5. Chen She
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  6. Hui Wang
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  7. Shuang Wu
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  8. Jianliang Zhang
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  9. Bing Zhu
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  10. Wanzhu Bai
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Contributions

This work presented here was carried out in collaboration between all authors. JW, DX, JC, CS, HW, SW, WB, SW, JZ and BZ carried out the experiments and analysed the data. JW and WB designed the study and wrote the paper. All authors read and approved the final version of the manuscript accepted for publication.

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Correspondence to Wanzhu Bai.

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Wang, J., Xu, D., Cui, J. et al. A new approach for examining the neurovascular structure with phalloidin and calcitonin gene-related peptide in the rat cranial dura mater. J Mol Hist 51, 541–548 (2020). https://doi.org/10.1007/s10735-020-09903-7

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  • DOI: https://doi.org/10.1007/s10735-020-09903-7

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