Overexpression of netrin-1 increases the expression of tight junction-associated proteins, claudin-5, occludin, and ZO-1, following traumatic brain injury in rats

Wen, Jianfeng; Suo-kai, Qian; Yang, Qifan; Deng, Lei; Mo, Ye; Yu, Yuefei

doi:10.3892/etm.2014.1818

Cited by 63 publications

(37 citation statements)

References 21 publications

(16 reference statements)

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“…Collected images were analyzed with ImageJ. Tight junction staining was conducted as detailed in [36], using ZO-1 and claudin-5 (Invitrogen, Grand Island, NY, USA) directly conjugated to Alexa 488 [37]. Briefly, greyscale measurements of the border between cell plasma membranes were taken for ten different cell-to-cell junctions in four different plates, and an average intensity was derived for each culture condition [23].…”

Section: Methodsmentioning

confidence: 99%

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Brown

Codreanu

Shi

et al. 2016

J Neuroinflammation

139

128

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BackgroundUnderstanding blood-brain barrier responses to inflammatory stimulation (such as lipopolysaccharide mimicking a systemic infection or a cytokine cocktail that could be the result of local or systemic inflammation) is essential to understanding the effect of inflammatory stimulation on the brain. It is through the filter of the blood-brain barrier that the brain responds to outside influences, and the blood-brain barrier is a critical point of failure in neuroinflammation. It is important to note that this interaction is not a static response, but one that evolves over time. While current models have provided invaluable information regarding the interaction between cytokine stimulation, the blood-brain barrier, and the brain, these approaches—whether in vivo or in vitro—have often been only snapshots of this complex web of interactions.MethodsWe utilize new advances in microfluidics, organs-on-chips, and metabolomics to examine the complex relationship of inflammation and its effects on blood-brain barrier function ex vivo and the metabolic consequences of these responses and repair mechanisms. In this study, we pair a novel dual-chamber, organ-on-chip microfluidic device, the NeuroVascular Unit, with small-volume cytokine detection and mass spectrometry analysis to investigate how the blood-brain barrier responds to two different but overlapping drivers of neuroinflammation, lipopolysaccharide and a cytokine cocktail of IL-1β, TNF-α, and MCP1,2.ResultsIn this study, we show that (1) during initial exposure to lipopolysaccharide, the blood-brain barrier is compromised as expected, with increased diffusion and reduced presence of tight junctions, but that over time, the barrier is capable of at least partial recovery; (2) a cytokine cocktail also contributes to a loss of barrier function; (3) from this time-dependent cytokine activation, metabolic signature profiles can be obtained for both the brain and vascular sides of the blood-brain barrier model; and (4) collectively, we can use metabolite analysis to identify critical pathways in inflammatory response.ConclusionsTaken together, these findings present new data that allow us to study the initial effects of inflammatory stimulation on blood-brain barrier disruption, cytokine activation, and metabolic pathway changes that drive the response and recovery of the barrier during continued inflammatory exposure.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-016-0760-y) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Brown

Codreanu

Shi

et al. 2016

J Neuroinflammation

139

128

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“…ZO-1 has been proven to regulate gap junction plaque size in various cell types, such as cardiomyocytes and neurons [37]. Additionally, occludin plays an important role in maintaining barrier function and TJ assembly [6]. A relevant study revealed that claudin-2 deficient mice are defective in the renal proximal tubules [38].…”

Section: Discussionmentioning

confidence: 99%

“…The IMB is mainly formed by the tight junctions of epithelial cells. Tight junctions consist of two proteins: the transmembrane proteins (occludins and claudins‐5), which can interact with adjacent endothelial cells to make up a physical barrier against paracellular diffusion, and the accessory proteins (Zonula occludens [ZO] family), which anchor the transmembrane proteins to the cytoskeleton [6]. ZO‐1 and occludin form a high resistance barrier in the bladder epithelium, and deterioration of these proteins marks the disruption of the pulmonary epithelial barrier [7–10].…”

Section: Introductionmentioning

confidence: 99%

Brain‐derived neurotrophic factor preserves intestinal mucosal barrier function and alters gut microbiota in mice

Cai

Zhang

2018

The Kaohsiung J of Med Scie

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The intestinal mucosal barrier (IMB) enables the intestine to provide adequate containment of luminal microorganisms and molecules while preserving the ability to absorb nutrients. In this study, we explored the effect of brain-derived neurotrophic factor (BDNF) on IMB function and gut microbiota in mice. BDNF gene knock-out mice (the BDNF group) and wild-type mice (the BDNF group) were selected. The gut microbiota of these mice was analyzed by denaturing gradient gel electrophoresis (DGGE) assay. The ultrastructure of the ileum and the colonic epithelium obtained from decapitated mice were observed by transmission electron microscopy. The protein expression of epithelial tight junction proteins, zonula occludens-1 (ZO-1) and occludin was detected by immunohistochemistry staining. The protein expression of claudin-1 and claudin-2 was determined by Western blotting. The DGGE band patterns of gut microbiota in the BDNF group were significantly different from that in the BDNF group, which indicated that the BDNF expression alters the gut microbiota in mice. Compared with the BDNF group, the BDNF group presented no significant difference in the ultrastructure of ileal epithelium; however, a significant difference was observed in the colonic epithelial barrier, manifested by decreased microvilli, widening intercellular space and bacterial invasion. Compared with the BDNF group, the expression of ZO-1 and occludin in the BDNF group was significantly decreased. The expression of claudin-1 in the BDNF group was significantly reduced, while the expression of claudin-2 was elevated. These findings indicate that BDNF preserves IMB function and modulates gut microbiota in mice.

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“…This result is very unlikely given the plethora of For (b, c), 232 ≤ N ≤ 244, where N is the number of cells, and for (d, e), N = 12, where N is the number of inserts pooled between the 0d and 1d cAMP conditions. The Mann-Whitney test was used to calculate significant differences for each parameter, where ns = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 literary evidence suggesting otherwise [38][39][40][41][42]. Importantly, however, many of these reports are qualitatively correlative between immunostaining and permeability measurement, and not a quantitative correlation between permeability and junction presentation.…”

Section: Transwell Permeability (And Teer) Assays Are Insufficient Fomentioning

confidence: 99%

Quantitatively relating brain endothelial cell–cell junction phenotype to global and local barrier properties under varied culture conditions via the Junction Analyzer Program

Gray

Jung

Inglut

et al. 2020

Fluids Barriers CNS

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Background: The endothelial cell-cell junctions of the blood-brain barrier (BBB) play a pivotal role in the barrier's function. Altered cell-cell junctions can lead to barrier dysfunction and have been implicated in several diseases. Despite this, the driving forces regulating junctional protein presentation remain relatively understudied, largely due to the lack of efficient techniques to quantify their presentation at sites of cell-cell adhesion. Here, we used our novel Junction Analyzer Program (JAnaP) to quantify junction phenotype (i.e., continuous, punctate, or perpendicular) in response to various substrate compositions, cell culture times, and cAMP treatments in human brain microvascular endothelial cells (HBMECs). We then quantitatively correlated junction presentation with barrier permeability on both a "global" and "local" scale. Methods:We cultured HBMECs on collagen I, fibronectin, collagen IV, laminin, fibronectin/collagen IV/laminin, or hyaluronic acid/gelatin for 2, 4, and 7 days with varying cAMP treatment schedules. Images of immunostained ZO-1, VE-cadherin, and claudin-5 were analyzed using the JAnaP to calculate the percent of the cell perimeter presenting continuous, punctate, or perpendicular junctions. Transwell permeability assays and resistance measurements were used to measure bulk ("global") barrier properties, and a "local" permeability assay was used to correlate junction presentation proximal to permeable monolayer regions.Results: Substrate composition was found to play little role in junction presentation, while cAMP supplements significantly increased the continuous junction architecture. Increased culture time required increased cAMP treatment time to reach similar ZO-1 and VE-cadherin coverage observed with shorter culture, though longer cultures were required for claudin-5 presentation. Prolonged cAMP treatment (6 days) disrupted junction integrity for all three junction proteins. Transwell permeability and TEER assays showed no correlation with junction phenotype, but a local permeability assay revealed a correlation between the number of discontinuous and no junction regions with barrier penetration. Conclusions:These results suggest that cAMP signaling influences HBMEC junction architecture more than matrix composition. Our studies emphasized the need for local barrier measurement to mechanistically understand the role of junction phenotype and supported previous results that continuous junctions are indicative of a more mature/ © The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article' s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material...

show abstract

Overexpression of netrin-1 increases the expression of tight junction-associated proteins, claudin-5, occludin, and ZO-1, following traumatic brain injury in rats

Cited by 63 publications

References 21 publications

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Brain‐derived neurotrophic factor preserves intestinal mucosal barrier function and alters gut microbiota in mice

Quantitatively relating brain endothelial cell–cell junction phenotype to global and local barrier properties under varied culture conditions via the Junction Analyzer Program

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