PUBLICATION

Intraperitoneal dye injection method for visualizing the functioning lymphatic vascular system in zebrafish and medaka

Authors
Saito, E., Isogai, S., Deguchi, T., Ishida, K., Nozaki, T., Ishiyama, E., Wayama, M., Shimoda, H.
ID
ZDB-PUB-191215-6
Date
2019
Source
Developmental Dynamics : an official publication of the American Association of Anatomists   249(5): 679-692 (Journal)
Registered Authors
Isogai, Sumio
Keywords
Cinnabar, Endothelial labeling, Lymphatic visualization, Trypan blue
MeSH Terms
  • Animals
  • Injections, Intraperitoneal
  • Lymphangiogenesis
  • Lymphatic Vessels*
  • Oryzias*
  • Reproducibility of Results
  • Zebrafish/genetics
PubMed
31837055 Full text @ Dev. Dyn.
Abstract
A hierarchically organized lymphatic vascular system extends throughout the vertebrate body for tissue fluid homeostasis, immune trafficking, and the absorption of dietary fats. Intralymphatic dye injection and serial sectioning have been the main tools for visualizing lymphatic vessels. Specific markers for identifying the lymphatic vasculature in zebrafish and medaka have appeared as new tools that enable the study of lymphangiogenesis using genetic and experimental manipulation. Transgenic fishes have become excellent organisms for visualizing the lymphatic vasculature in living embryos, but this method has limited usefulness, especially in later developmental stages. The functional lymphatic endothelium predominantly takes up foreign particles in zebrafish and medaka. We utilized this physiological activity and lymph flow to label lymphatic vessels. Intraperitoneal injection of trypan blue is useful for temporal observations of the lymphatic ducts, which are essential for large-scale genetic screening, while cinnabar (HgS) injection allows identification of the lymphatic endothelium under electron microscopy, avoiding artifactual damage. This injection method, which is not high in cost and does not require high skill or special devices, is applicable to any live fish with functioning lymphatic vessels, even mutants, with high reproducibility for visualizing the entire lymphatic vascular system. This article is protected by copyright. All rights reserved.
Genes / Markers
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Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping