Correlation of zonula occludens ZO-1 antigen expression and transendothelial resistance in porcine and rat cultured cerebral endothelial cells

Krause, Dorothee; Mischeck, Uwe; Galla, Hans‐Joachim; Dermietzel, Rolf

doi:10.1016/0304-3940(91)90284-z

Cited by 43 publications

(20 citation statements)

References 17 publications

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“…The detection of the tight junction marker Zonula occludens (ZO-1) (even in the absence of astrocytes) ( Fig. 1A and B), by staining directly adherent BMEC monolayers with a specific antibody, suggests that the isolated BMECs used in this study represent appropriate targets for HIV-1 transmigration analysis (42,74). Consistent with the in vitro immunostaining data, we found that human brain capillaries also express high levels of heparan sulfates in vivo (Fig.…”

Section: Cell Surface Expression Of Proteoglycans On Primary Humansupporting

confidence: 83%

Contribution of Proteoglycans to Human Immunodeficiency Virus Type 1 Brain Invasion

Bobardt

Salmon

Wang

et al. 2004

J Virol

106

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As a neurotropic virus, human immunodeficiency virus type 1 (HIV-1) invades the brain and causes severe neuronal, astrocyte, and myelin damage in AIDS patients. To gain access to the brain, HIV-1 must migrate through brain microvascular endothelial cells (BMECs), which compose the blood-brain barrier (BBB). Given that BMECs lack the entry receptor CD4, HIV-1 must use receptors distinct from CD4 to enter these cells. We previously reported that cell surface proteoglycans serve as major HIV-1 receptors on primary human endothelial cells. In this study, we examined whether proteoglycans also impact cell-free HIV-1 invasion of the brain. Using an artificial BBB transmigration assay, we found that both heparan and chondroitin sulfate proteoglycans (HSPGs and CSPGs, respectively) are abundantly expressed on primary BMECs and promote HIV-1 attachment and entry. In contrast, the classical entry receptors, CXCR4 and CCR5, only moderately enhanced these processes. HSPGs and CSPGs captured HIV-1 in a gp120-dependent manner. However, no correlation between coreceptor usage and transmigration was identified. Furthermore, brain-derived viruses did not transmigrate more efficiently than lymphoid-derived viruses, suggesting that the ability of HIV-1 to replicate in the brain does not correlate with its capacity to migrate through the BBB as cell-free virus. Given that HIV-1-proteoglycan interactions are based on electrostatic contacts between basic residues in gp120 and sulfate groups in proteoglycans, HIV-1 may exploit these interactions to rapidly enter and migrate through the BBB to invade the brain.

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Section: Cell Surface Expression Of Proteoglycans On Primary Humansupporting

confidence: 83%

Contribution of Proteoglycans to Human Immunodeficiency Virus Type 1 Brain Invasion

Bobardt

Salmon

Wang

et al. 2004

J Virol

106

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“…TEER in mouse LMECs monolayer on day 6 after inoculation increased up to approximately 19.0 ± 2.9 Ω × cm 2 . This value was similar to that of cultured wild-type rat lung endothelial cells [10], but was approximately 20% lower than that of cultured rat brain endothelial cells in the confluent condition [11] [12]. On the other hand, the paracellular permeability of mouse LMEC monolayers by the water soluble small marker fluorescein is the lowest on day 6 after inoculation.…”

Section: Discussionsupporting

confidence: 63%

“…Moreover, several peripheral membrane proteins, such as ZO-1 [23], ZO-2 [24], ZO-3 [18], cingulin [25], 7H6 antigen [26] and symplekin [27] were shown to be concentrated at the cytoplasmic surface of TJs. Freshly isolated cerebral endothelial cell culture aggregates and 7-day-old cerebral endothelial cell culture in primary culture express ZO-1 protein [11]. In this study, the expression of TJ-associated proteins occludin, claudin-1, -4 and ZO-1 was observed in LMECs on day 6 in culture when paracellular permeability was the lowest.…”

Section: Discussionmentioning

confidence: 49%

Claudin-1 Leads to Strong Formation of Tight Junction in Cultured Mouse Lung Microvascular Endothelial Cells

Ueda¹,

Shinmyouzu²,

Nakayama³

et al. 2016

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We aimed to examine paracellular barrier function in cultured mouse lung microvascular endothelial cells (LMECs). The transcellular resistance of LMEC monolayers yielded an electrical resistance of approximately 19 Ω × cm 2 at days 6 -7 in culture when the cells reached confluence, and paracellular permeable clearance of sodium fluorescein was the lowest on day 6 in culture, suggesting the formation of tight junctions (TJs) in cultured LMECs. Moreover, the expression of TJassociated proteins, occludin, claudin-1 and -4 and zonula occludents 1 (ZO-1) was detected in LMECs at day 6 in culture. However, mRNAs of occludin, claudin-1 and -4 and ZO-1 were already expressed on day 1 after culture, and large variations were absent in the mRNA levels of occludin, claudin-4 and ZO-1 between days 1 and 7 in culture, when the level of each mRNA on day 1 in culture was used as a basal level. However, the claudin-1 mRNA level gradually increased up to approximately 7-fold on day 7 in culture over the basal level. These results indicate that the drastic increase in the mRNA expression level of claudin-1 leads to the strong formation of TJs.

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“…ZO-1 expression at the cell borders for iPSC-ECs (Fig. 2A) provided evidence of tight junction formation[36, 37]. We further investigated the ability of iPSC-ECs to form a barrier using an impedance-based platform to assess barrier function (see Methods)[34].…”

Section: Resultsmentioning

confidence: 99%

Human Vascular Tissue Models Formed from Human Induced Pluripotent Stem Cell Derived Endothelial Cells

Belair

Whisler

Valdez

et al. 2014

Stem Cell Rev and Rep

112

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Here we describe a strategy to model blood vessel development using a well-defined iPSC-derived endothelial cell type (iPSC-EC) cultured within engineered platforms that mimic the 3D microenvironment. The iPSC-ECs used here were first characterized by expression of endothelial markers and functional properties that included VEGF responsiveness, TNF-α-induced upregulation of cell adhesion molecules (MCAM/CD146; ICAM1/CD54), thrombin-dependent barrier function, shear stress-induced alignment, and 2D and 3D capillary-like network formation in Matrigel. The iPSC-ECs also formed 3D vascular networks in a variety of engineering contexts, yielded perfusable, interconnected lumen when co-cultured with primary human fibroblasts, and aligned with flow in microfluidics devices. iPSC-EC function during tubule network formation, barrier formation, and sprouting was consistent with that of primary ECs, and the results suggest a VEGF-independent mechanism for sprouting, which is relevant to therapeutic anti-angiogenesis strategies. Our combined results demonstrate the feasibility of using a well-defined, stable source of iPSC-ECs to model blood vessel formation within a variety of contexts using standard in vitro formats.

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Correlation of zonula occludens ZO-1 antigen expression and transendothelial resistance in porcine and rat cultured cerebral endothelial cells

Cited by 43 publications

References 17 publications

Contribution of Proteoglycans to Human Immunodeficiency Virus Type 1 Brain Invasion

Contribution of Proteoglycans to Human Immunodeficiency Virus Type 1 Brain Invasion

Claudin-1 Leads to Strong Formation of Tight Junction in Cultured Mouse Lung Microvascular Endothelial Cells

Human Vascular Tissue Models Formed from Human Induced Pluripotent Stem Cell Derived Endothelial Cells

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