Visualization and Analysis of Pharyngeal Arch Arteries using Whole-mount Immunohistochemistry and 3D Reconstruction.
Journal
Journal of visualized experiments : JoVE
ISSN: 1940-087X
Titre abrégé: J Vis Exp
Pays: United States
ID NLM: 101313252
Informations de publication
Date de publication:
31 03 2020
31 03 2020
Historique:
entrez:
21
4
2020
pubmed:
21
4
2020
medline:
2
10
2020
Statut:
epublish
Résumé
Improper formation or remodeling of the pharyngeal arch arteries (PAAs) 3, 4, and 6 contribute to some of the most severe forms of congenital heart disease. To study the formation of PAAs, we developed a protocol using whole-mount immunofluorescence coupled with benzyl alcohol/benzyl benzoate (BABB) tissue clearing, and confocal microscopy. This allows for the visualization of the pharyngeal arch endothelium at a fine cellular resolution as well as the 3D connectivity of the vasculature. Using software, we have established a protocol to quantify the number of endothelial cells (ECs) in PAAs, as well as the number of ECs within the vascular plexus surrounding the PAAs within pharyngeal arches 3, 4, and 6. When applied to the whole embryo, this methodology provides a comprehensive visualization and quantitative analysis of embryonic vasculature.
Identifiants
pubmed: 32310236
doi: 10.3791/60797
pmc: PMC7216781
mid: NIHMS1585261
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Video-Audio Media
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NHLBI NIH HHS
ID : F31 HL150949
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL103920
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL134935
Pays : United States
Organisme : NIH HHS
ID : R21 OD025323
Pays : United States
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