Studying the Neurovascular Unit: An Improved Blood–Brain Barrier Model

Zehendner, Christoph M.; Luhmann, Heiko J.; Kuhlmann, Christoph

doi:10.1038/jcbfm.2009.103

Cited by 23 publications

(29 citation statements)

References 14 publications

(16 reference statements)

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“…We used a murine brain endothelial cell line bEnd.3 (bEnd.3, American Type Culture Collection, Manassas, Virginia, USA) for coculture experiments as described previously [15]. bEnd.3 were cultured as recommended by the manufacturer and maintained at 37°C and 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…We used a recently described model of the NVU [15] to analyze (i) acute ischemic alterations of the NVU by immunohistochemistry of the TJ proteins Zonula occludens-1 (ZO-1) and Claudin-5 (Cl-5), which have a key role in the pathogenesis of ischemic blood-brain barrier breakdown [6], [15]–[17], and (ii) whether caspase-3 is involved in altering TJ protein arrangement. To strengthen the significance of our investigation we also performed immunohistochemical analyzes of ZO-1 after middle cerebral artery occlusion (MCAO) in the guinea pig isolated whole brain, in which the BBB as NVU is completely preserved [18].…”

Section: Introductionmentioning

confidence: 99%

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Caspase-3 Contributes to ZO-1 and Cl-5 Tight-Junction Disruption in Rapid Anoxic Neurovascular Unit Damage

et al. 2011

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BackgroundTight-junction (TJ) protein degradation is a decisive step in hypoxic blood-brain barrier (BBB) breakdown in stroke. In this study we elucidated the impact of acute cerebral ischemia on TJ protein arrangement and the role of the apoptotic effector protease caspase-3 in this context.Methodology/Principal FindingsWe used an in vitro model of the neurovascular unit and the guinea pig whole brain preparation to analyze with immunohistochemical methods the BBB properties and neurovascular integrity. In both methodological approaches we observed rapid TJ protein disruptions after 30 min of oxygen and glucose deprivation or middle cerebral artery occlusion, which were accompanied by strong caspase-3 activation in brain endothelial cells (BEC). Surprisingly only few DNA-fragmentations were detected with TUNEL stainings in BEC. Z-DEVD-fmk, an irreversible caspase-3 inhibitor, partly blocked TJ disruptions and was protective on trans-endothelial electrical resistance.Conclusions/SignificanceOur data provide evidence that caspase-3 is rapidly activated during acute cerebral ischemia predominantly without triggering DNA-fragmentation in BEC. Further we detected fast TJ protein disruptions which could be partly blocked by caspase-3 inhibition with Z-DEVD-fmk. We suggest that the basis for clinically relevant BBB breakdown in form of TJ disruptions is initiated within minutes during ischemia and that caspase-3 contributes to this process.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Caspase-3 Contributes to ZO-1 and Cl-5 Tight-Junction Disruption in Rapid Anoxic Neurovascular Unit Damage

et al. 2011

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“…4b). It is known that metabolic inhibition with the mitochondrial respiratory chain inhibitor sodium cyanide (NaCN) closely mimics the features of hypoxia at the blood brain barrier [21]. PDTC …”

Section: Pdtc Inhibits Apoptotic Cell Death Of Cerebral Microvascularmentioning

confidence: 99%

Pyrrolidine Dithiocarbamate Activates p38 MAPK and Protects Brain Endothelial Cells From Apoptosis: A Mechanism for the Protective Effect in Stroke?

et al. 2010

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The antioxidant and inhibitor of nuclear factor kappaB pyrrolidine dithiocarbamate (PDTC) potently reduces infarct size in various experimental stroke models. In addition, it has been shown to have a favourable safety profile in humans. In this study, we further investigated the mechanistic actions of PDTC on cerebral microvascular endothelial cells as main components of the blood-brain barrier. We propose activation of p38 MAPK by PDTC as an additional protective mechanism. C57/BL6 mice were subjected to transient MCAO for 2 h and treated with PDTC (100 mg/kg) or vehicle i.p. before reperfusion. Infarct size was determined after 24 h. Apoptosis was induced in a cerebral microvascular endothelial cell line and the effect of pretreatment with PDTC and its dependency on p38 MAPK activity was assayed. PDTC administered 2 h after MCAO reduced infarct size by 61% (P < 0.05) and reduces the apoptotic death rate in vivo. In vitro, PDTC reduces the apoptotic death rate of bEnd.3 cells. p38 MAPK was activated by PDTC and its inhibition abrogated the protective effect of PDTC. PDTC reduces infarct size after stroke with a reasonable time window and decreases apoptotic cell death in vivo and in vitro. The attenuation of apoptotic cell death in brain microvascular endothelial cells is dependent on p38 MAPK activity.

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“…The different combination of cerebral endothelial cells with elements of the neurovascular unit makes coculture systems differ from each other [288][289][290]. On the one hand, cerebral endothelial cells can be established with either primary cells or cell lines, while glial cells (which can be primary astrocytes, astrocytic cell lines or C6 glioma cell lines), pericytes or even neurons can be included as elements of the neurovascular unit.…”

Section: Coculture Systemsmentioning

confidence: 99%

In vitro screening of nanomedicines through the blood brain barrier: A critical review

2016

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Studying the Neurovascular Unit: An Improved Blood–Brain Barrier Model

Cited by 23 publications

References 14 publications

Caspase-3 Contributes to ZO-1 and Cl-5 Tight-Junction Disruption in Rapid Anoxic Neurovascular Unit Damage

Caspase-3 Contributes to ZO-1 and Cl-5 Tight-Junction Disruption in Rapid Anoxic Neurovascular Unit Damage

Pyrrolidine Dithiocarbamate Activates p38 MAPK and Protects Brain Endothelial Cells From Apoptosis: A Mechanism for the Protective Effect in Stroke?

In vitro screening of nanomedicines through the blood brain barrier: A critical review

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