Fluorescent imaging of endothelial glycocalyx layer with wheat germ agglutinin using intravital microscopy

Microscopy Research and Technique
Hanae KataokaTakehiko Iijima

Abstract

Endothelial glycocalyx (GCX) is located on the apical surface of vascular endothelial cells and is composed of a negatively-charged network of proteoglycans and glycoproteins. The GCX plays an important role in maintaining the integrity of vascular walls and preventing leakage of plasma. Therefore, degradation of the GCX is believed to lead to pathological leakage of plasma. Because the GCX is a very thin layer, its ultrastructural image has been demonstrated on electron microscope. To explore the function of the GCX, it should be visualized by a microscope in vivo. Thus, we developed in vivo visualization technique of the GCX under fluorescence microscopy using a mouse dorsal skinfold chamber (DSC) model. To label and visualize the GCX, we used fluorescein isothiocyanate (FITC)-labeled lectin, which has a high specificity for sugar moieties. We examined the affinity of the different lectins to epivascular regions under an intravital fluorescent microscope. Among seven different lectins we examined, FITC labeled Triticum vulgaris (wheat germ) agglutinin (WGA) delineated the GCX most clearly. Binding of WGA to the GCX was inhibited by chitin hydrolysate, which contained WGA-binding polysaccharide chains. Furthermore, the septic ...Continue Reading

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Citations

Sep 13, 2016·Journal of Intensive Care·Akira UshiyamaTakehiko Iijima
Jan 12, 2019·American Journal of Physiology. Heart and Circulatory Physiology·Mohamed M AliShane A Phillips
Feb 19, 2020·Journal of Veterinary Emergency and Critical Care·Sarah GaudetteManuel Boller
Oct 19, 2017·The Journal of Trauma and Acute Care Surgery·Mark E DiebelLawrence N Diebel
Sep 24, 2019·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Tian NiuKun Liu
Dec 7, 2017·Microcirculation : the Official Journal of the Microcirculatory Society, Inc·Petra NijstWilfried Mullens
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Apr 9, 2019·Journal of the Royal Society, Interface·Viola IntroiniStefano Guido
Nov 2, 2019·Investigative Ophthalmology & Visual Science·Kiumars ShamlooAjay Sharma
Feb 10, 2020·The American Journal of Pathology·Sophie Dogné, Bruno Flamion
Mar 7, 2020·Biophysical Journal·Luis F DelgadilloRichard E Waugh
Jan 28, 2021·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Kyle H MooreEric M George
Jun 1, 2021·Frontiers in Veterinary Science·Edward S Cooper, Deborah C Silverstein
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Jun 12, 2021·The American Journal of Pathology·Haruka OkamotoShinji Ogura
Jun 16, 2021·Translational Vision Science & Technology·Kiumars ShamlooAjay Sharma

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