title: Three-Dimensional Visualization of Developing Neurovascular Architecture in the Craniofacial Region of Embryonic Mice.
abstract: 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.
authors.affiliation: Department of Pathology, The Nippon Dental University School of Life Dentistry at Tokyo, Chiyoda-ku, Tokyo, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Sugimoto
authors.affiliation: Department of Pathology, The Nippon Dental University School of Life Dentistry at Tokyo, Chiyoda-ku, Tokyo, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Taya
authors.affiliation: Department of Food and Life Science, School of Life and Environmental Science, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Shimazu
authors.affiliation: Department of Pathology, The Nippon Dental University School of Life Dentistry at Tokyo, Chiyoda-ku, Tokyo, Japan.
authors.forename: Yuuichi
authors.initials: Y
authors.lastname: Soeno
authors.affiliation: Department of Pathology, The Nippon Dental University School of Life Dentistry at Tokyo, Chiyoda-ku, Tokyo, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Sato
authors.affiliation: Department of Pathology, The Nippon Dental University School of Life Dentistry at Tokyo, Chiyoda-ku, Tokyo, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Aoba
mesh_terms: D000818:Animals; D001369:Axons; D001808:Blood Vessels / Q000033:anatomy & histology*; D003391:Cranial Nerves / Q000033:anatomy & histology* / Q000196:embryology; D005145:Face / Q000033:anatomy & histology* / Q000196:embryology; D005260:Female; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008813:Mice, Inbred ICR; D009420:Nervous System / Q000033:anatomy & histology*; D011247:Pregnancy
keywords: axon growth; cranial nerve; craniofacial development; embryonic mouse; neurovascular patterning; three-dimensional reconstruction; vascular remodeling