Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain

Tight homeostatic control of brain amino acids (AA) depends on transport by solute carrier family proteins expressed by the blood—brain barrier (BBB) microvascular endothelial cells (BMEC). To characterize the mouse BMEC transcriptome and probe culture-induced changes, microarray analyses of platelet endothelial cell adhesion molecule-1-positive (PECAM1+) endothelial cells (ppMBMECs) were compared with primary MBMECs (pMBMEC) cultured in the presence or absence of glial cells and with b.End5 endothelioma cell line. Selected cell marker and AA transporter mRNA levels were further verified by reverse transcription real-time PCR. Regardless of glial coculture, expression of a large subset of genes was strongly altered by a brief culture step. This is consistent with the known dependence of BMECs on in vivo interactions to maintain physiologic functions, for example, tight barrier formation, and their consequent dedifferentiation in culture. Seven ( 4F2hc, Lat1, Taut, Snat3, Snat5, Xpct, and Cat1) of nine AA transporter mRNAs highly expressed in freshly isolated ppMBMECs were strongly downregulated for all cultures and two ( Snat2 and Eaat3) were variably regulated. In contrast, five AA transporter mRNAs with low expression in ppMBMECs, including y+ Lat2, xCT, and Snat1, were upregulated by culture. We hypothesized that the AA transporters highly expressed in ppMBMECs and downregulated in culture have a major in vivo function for BBB transendothelial transport.

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Podocalyxin is required for maintaining blood–brain barrier function during acute inflammation

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1814766116 ◽

2019 ◽

Vol 116 (10) ◽

pp. 4518-4527 ◽

Cited By ~ 8

Author(s):

Jessica Cait ◽

Michael R. Hughes ◽

Matthew R. Zeglinski ◽

Allen W. Chan ◽

Sabrina Osterhof ◽

...

Keyword(s):

Endothelial Cells ◽

Blood Brain Barrier ◽

Barrier Function ◽

Vascular Endothelial Cells ◽

Umbilical Vein ◽

Focal Adhesions ◽

Brain Barrier ◽

Cortical Actin ◽

Inflammatory Conditions ◽

Blood Brain

Podocalyxin (Podxl) is broadly expressed on the luminal face of most blood vessels in adult vertebrates, yet its function on these cells is poorly defined. In the present study, we identified specific functions for Podxl in maintaining endothelial barrier function. Using electrical cell substrate impedance sensing and live imaging, we found that, in the absence of Podxl, human umbilical vein endothelial cells fail to form an efficient barrier when plated on several extracellular matrix substrates. In addition, these monolayers lack adherens junctions and focal adhesions and display a disorganized cortical actin cytoskeleton. Thus, Podxl has a key role in promoting the appropriate endothelial morphogenesis required to form functional barriers. This conclusion is further supported by analyses of mutant mice in which we conditionally deleted a floxed allele ofPodxlin vascular endothelial cells (vECs) using Tie2Cre mice (PodxlΔTie2Cre). Although we did not detect substantially altered permeability in naïve mice, systemic priming with lipopolysaccharide (LPS) selectively disrupted the blood–brain barrier (BBB) inPodxlΔTie2Cremice. To study the potential consequence of this BBB breach, we used a selective agonist (TFLLR-NH2) of the protease-activated receptor-1 (PAR-1), a thrombin receptor expressed by vECs, neuronal cells, and glial cells. In response to systemic administration of TFLLR-NH2, LPS-primedPodxlΔTie2Cremice become completely immobilized for a 5-min period, coinciding with severely dampened neuroelectric activity. We conclude that Podxl expression by CNS tissue vECs is essential for BBB maintenance under inflammatory conditions.

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Isoflurane Induces Endothelial Apoptosis of the Post-Hypoxic Blood-Brain Barrier in a Transdifferentiated Human Umbilical Vein Edothelial Cell Model

PLoS ONE ◽

10.1371/journal.pone.0038260 ◽

2012 ◽

Vol 7 (6) ◽

pp. e38260 ◽

Cited By ~ 13

Author(s):

Michael S. Dittmar ◽

Walter Petermichl ◽

Felix Schlachetzki ◽

Bernhard M. Graf ◽

Michael Gruber

Keyword(s):

Blood Brain Barrier ◽

Cell Model ◽

Umbilical Vein ◽

Brain Barrier ◽

Human Umbilical Vein ◽

Endothelial Apoptosis ◽

Blood Brain

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Mouse Adenovirus Type 1-Induced Breakdown of the Blood-Brain Barrier

Journal of Virology ◽

10.1128/jvi.00954-09 ◽

2009 ◽

Vol 83 (18) ◽

pp. 9398-9410 ◽

Cited By ~ 39

Author(s):

Lisa E. Gralinski ◽

Shanna L. Ashley ◽

Shandee D. Dixon ◽

Katherine R. Spindler

Keyword(s):

Endothelial Cells ◽

Blood Brain Barrier ◽

Mouse Brain ◽

Adenovirus Type ◽

Brain Barrier ◽

Brain Endothelial Cells ◽

Blood Brain ◽

Mouse Brain Endothelial Cells ◽

Primary Mouse ◽

Infected Cells

ABSTRACT Infection with mouse adenovirus type 1 (MAV-1) results in fatal acute encephalomyelitis in susceptible mouse strains via infection of brain endothelial cells. Wild-type (wt) MAV-1 causes less brain inflammation than an early region 3 (E3) null virus in C57BL/6 mice. A mouse brain microvascular endothelial cell line infected with wt MAV-1 had higher expression of mRNAs for the proinflammatory chemokines CCL2 and CCL5 than mock- and E3 null virus-infected cells. Primary mouse brain endothelial cells infected with wt virus had elevated levels of CCL2 compared to mock- or E3 null virus-infected cells. Infection of C57BL/6 mice with wt MAV-1 or the E3 null virus caused a dose-dependent breakdown of the blood-brain barrier, primarily due to direct effects of virus infection rather than inflammation. The tight junction proteins claudin-5 and occludin showed reduced surface expression on primary mouse brain endothelial cells following infection with either wt MAV-1 or the E3 null virus. mRNAs and protein for claudin-5, occludin, and zona occludens 2 were also reduced in infected cells. MAV-1 infection caused a loss of transendothelial electrical resistance in primary mouse brain endothelial cells that was not dependent on E3 or on MAV-1-induced CCL2 expression. Taken together, these results demonstrate that MAV-1 infection caused breakdown of the blood-brain barrier accompanied by decreased surface expression of tight junction proteins. Furthermore, while the MAV-1-induced pathogenesis and inflammation were dependent on E3, MAV-1-induced breakdown of the blood-brain barrier and alteration of endothelial cell function were not dependent on E3 or CCL2.

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Nitric oxide mediates cyclosporine-induced impairment of the blood–brain barrier in cocultures of mouse brain endothelial cells and rat astrocytes

European Journal of Pharmacology ◽

10.1016/j.ejphar.2004.10.027 ◽

2004 ◽

Vol 505 (1-3) ◽

pp. 51-59 ◽

Cited By ~ 14

Author(s):

Shinya Dohgu ◽

Atsushi Yamauchi ◽

Shinsuke Nakagawa ◽

Fuyuko Takata ◽

Mamiko Kai ◽

...

Keyword(s):

Nitric Oxide ◽

Endothelial Cells ◽

Blood Brain Barrier ◽

Mouse Brain ◽

Brain Barrier ◽

Brain Endothelial Cells ◽

Blood Brain ◽

Mouse Brain Endothelial Cells ◽

Rat Astrocytes

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Time-Dependent Internalization of Polymer-Coated Silica Nanoparticles in Brain Endothelial Cells and Morphological and Functional Effects on the Blood-Brain Barrier

International Journal of Molecular Sciences ◽

10.3390/ijms22041657 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1657

Author(s):

Aniela Bittner ◽

Fabien Gosselet ◽

Emmanuel Sevin ◽

Lucie Dehouck ◽

Angélique D. Ducray ◽

...

Keyword(s):

Endothelial Cells ◽

Blood Brain Barrier ◽

Cell Types ◽

Tube Formation ◽

Time Dependent ◽

Brain Barrier ◽

Brain Endothelial Cells ◽

Blood Brain ◽

Endothelial Tube Formation ◽

Barrier Formation

Nanoparticle (NP)-assisted procedures including laser tissue soldering (LTS) offer advantages compared to conventional microsuturing, especially in the brain. In this study, effects of polymer-coated silica NPs used in LTS were investigated in human brain endothelial cells (ECs) and blood-brain barrier models. In the co-culture setting with ECs and pericytes, only the cell type directly exposed to NPs displayed a time-dependent internalization. No transfer of NPs between the two cell types was observed. Cell viability was decreased relatively to NP exposure duration and concentration. Protein expression of the nuclear factor ĸ-light-chain-enhancer of activated B cells and various endothelial adhesion molecules indicated no initiation of inflammation or activation of ECs after NP exposure. Differentiation of CD34+ ECs into brain-like ECs co-cultured with pericytes, blood-brain barrier (BBB) characteristics were obtained. The established endothelial layer reduced the passage of integrity tracer molecules. NP exposure did not result in alterations of junctional proteins, BBB formation or its integrity. In a 3-dimensional setup with an endothelial tube formation and tight junctions, barrier formation was not disrupted by the NPs and NPs do not seem to cross the blood-brain barrier. Our findings suggest that these polymer-coated silica NPs do not damage the BBB.

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