Yamazaki Tomoko, Li Wenling, Mukouyama Yoh-Suke
Laboratory of Stem Cell and Neuro-Vascular Biology, Genetics and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health; Earle A. Chiles Research Institute, Robert W. Franz Cancer Center, Providence Portland Medical Center.
Laboratory of Stem Cell and Neuro-Vascular Biology, Genetics and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health.
J Vis Exp. 2018 Mar 29(133):57406. doi: 10.3791/57406.
Here, we present a protocol of a whole-mount adult ear skin imaging technique to study comprehensive three-dimensional neuro-vascular branching morphogenesis and patterning, as well as immune cell distribution at a cellular level. The analysis of peripheral nerve and blood vessel anatomical structures in adult tissues provides some insights into the understanding of functional neuro-vascular wiring and neuro-vascular degeneration in pathological conditions such as wound healing. As a highly informative model system, we have focused our studies on adult ear skin, which is readily accessible for dissection. Our simple and reproducible protocol provides an accurate depiction of the cellular components in the entire skin, such as peripheral nerves (sensory axons, sympathetic axons, and Schwann cells), blood vessels (endothelial cells and vascular smooth muscle cells), and inflammatory cells. We believe this protocol will pave the way to investigate morphological abnormalities in peripheral nerves and blood vessels as well as the inflammation in the adult ear skin under different pathological conditions.
在此，我们展示了一种成年耳部皮肤整体成像技术的方案，用于在细胞水平研究全面的三维神经血管分支形态发生和模式形成，以及免疫细胞分布。对成年组织中周围神经和血管解剖结构的分析为理解诸如伤口愈合等病理状况下的功能性神经血管布线和神经血管退化提供了一些见解。作为一个信息丰富的模型系统，我们将研究重点放在了成年耳部皮肤上，它易于进行解剖。我们简单且可重复的方案能够准确描绘整个皮肤中的细胞成分，如周围神经（感觉轴突、交感轴突和施万细胞）、血管（内皮细胞和血管平滑肌细胞）以及炎症细胞。我们相信该方案将为研究不同病理状况下成年耳部皮肤中周围神经和血管的形态异常以及炎症铺平道路。
