Three-dimensional visualization of developing neurovascular architecture in the craniofacial region of embryonic mice
Three-dimensional visualization of developing neurovascular architecture in the craniofacial region of embryonic mice
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胚胎小鼠颅面部区域神经血管结构发育的三维可视化
DOI：
10.1002/ar.23179
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Aoba T
中科院分区：
文献类型：
--
作者：
Sugimoto T;Taya Y;Shimazu Y;Soeno Y;Sato K;Aoba T
最近的研究强调了血管和轴突引导的机制，以确保适当的形态发生和器官发生。我们的目的是对胚胎小鼠颅面发育过程中神经血管网络的发育进行全局绘图。为此，我们使用从小鼠胚胎获得的石蜡包埋的连续切片开发了基于组织学的三维（3D）重建。所有连续切片均使用 Pecam1 和 Pgp9.5/Gap43 混合抗体进行双重免疫标记。所有免疫标记的连续切片均通过虚拟显微镜进行数字化，以获得高空间分辨率图像。 3D 重建保证了卓越的位置精度，以追踪血管和单个颅神经轴突的远程连接。同时描绘血管生成萌芽和轴突末端树枝化的细节并定量评估血管和颅神经轴突之间的局部区域接近度是可行的。值得注意的是，颅面部区域的 3D 视图揭示了以下内容：在胚胎 (E) 9.5 天，鳃弓动脉和毛细血管丛在没有伴随神经的情况下形成。颅神经轴突开始长入鳃弓，在 E10.5 处形成小血管迷宫。发生血管重塑，上颌骨、下颌骨、鼓索和舌下神经轴突的轴突末端在 E11.5 的外侧舌肿胀周围形成树枝状。三叉神经和颈动脉分支的分歧模式在 E11.5 处变得一致。总体结果支持双免疫标记和 3D 重建技术在记录小鼠胚胎中发育中的神经血管网络的架构和布线方面的优势。 Anat Rec，298：1824–1835，2015。© 2015 Wiley periodicals, Inc.
Recent studies have highlighted the mechanism of vascular and axonal guidance to ensure proper morphogenesis and organogenesis. We aimed to perform global mapping of developing neurovascular networks during craniofacial development of embryonic mice. To this end, we developed histology‐based three‐dimensional (3D) reconstructions using paraffin‐embedded serial sections obtained from mouse embryos. All serial sections were dual‐immunolabeled with Pecam1 and Pgp9.5/Gap43 cocktail antibodies. All immunolabeled serial sections were digitized with virtual microscopy to acquire high spatial resolution images. The 3D reconstructs warranted superior positional accuracy to trace the long‐range connectivity of blood vessels and individual cranial nerve axons. It was feasible to depict simultaneously the details of angiogenic sprouting and axon terminal arborization and to assess quantitatively the locoregional proximity between blood vessels and cranial nerve axons. Notably, 3D views of the craniofacial region revealed the following: Branchial arch arteries and blood capillary plexi were formed without accompanying nerves at embryonic day (E) 9.5. Cranial nerve axons began to grow into the branchial arches, developing a labyrinth of small blood vessels at E10.5. Vascular remodeling occurred, and axon terminals of the maxillary, mandibular, chorda tympani, and hypoglossal nerve axons had arborized around the lateral lingual swellings at E11.5. The diverged patterning of trigeminal nerves and the arterial branches from the carotid artery became congruent at E11.5. The overall results support the advantage of dual‐immunolabeling and 3D reconstruction technology to document the architecture and wiring of the developing neurovascular networks in mouse embryos. Anat Rec, 298:1824–1835, 2015. © 2015 Wiley Periodicals, Inc.