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Updated: Aug 2, 2026

Isolation of Primary Murine Brain Microvascular Endothelial Cells
Published on: November 14, 2014
Immunocytochemical expression of the endothelial barrier antigen (EBA) during brain angiogenesis
J M Rosenstein1, J M Krum, L A Sternberger
1Department of Anatomy, George Washington University Medical Center, Washington, DC 20037.
Abstract:
The antibody to the endothelial barrier antigen (anti-EBA) is localized to the luminal plasma membrane of endothelia that have a blood-brain barrier (BBB) but not to other vessels, for instance those in the circumventricular organs, which lack barrier function. We have examined EBA expression in the rat in certain tissues and in brain microvessels in models of brain angiogenesis such as development, wound healing and neural transplantation. All brain microvessels including pial ones stained for anti-EBA whereas those of the dura, median eminence and choroid plexus did not. Vessels of the iris which are characterized by tight junctions and barrier function expressed EBA strongly. Embryonic day 18 brain did not stain at all for anti-EBA although vessels were readily localized with anti-laminin. Following stab wounds to mature brain, directly injured and adjacent microvessels lacked EBA expression for a period of approximately 2 weeks which is a similar time frame of BBB breakdown. Following this period, EBA expression gradually returned to a normal pattern by 3-4 weeks. Likewise, in intraparenchymal transplants of fetal neocortex EBA expression was not observed for 2 weeks and while at later times transplant vessels expressed EBA whereas some interface vessels associated with inflammatory cells did not. Permeable choroid plexus vessels vascularizing intraventricular transplants did not stain for anti-EBA at any time period and neither did vessels in adrenal medulla transplants. The present study shows that while EBA expression is a postnatal event unlike the development of a barrier to serum protein, its expression may be lost or delayed in injured vessels or ones associated with inflammatory cells or reactive astrocytes.
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