scholarly journals The Use of Microdialysis Techniques in Mice to Study P-gp Function at the Blood-Brain Barrier

2012 ◽  
Vol 18 (4) ◽  
pp. 430-440 ◽  
Author(s):  
István Sziráki ◽  
Franciska Erdő ◽  
Péter Trampus ◽  
Mirabella Sike ◽  
Petra Magdolna Molnár ◽  
...  
Keyword(s):  
Drug Interactions ◽  
Blood Brain Barrier ◽  
Test System ◽  
Assay System ◽  
Brain Barrier ◽  
Psc 833 ◽  
Blood Brain ◽  
Primary Mouse ◽  
Substrate Inhibitor ◽  
Inhibitor Combination

An integrated assay system involving dual/triple-probe microdialysis techniques in rats was developed earlier for testing interactions with P-glycoprotein (P-gp) at the blood-brain barrier using quinidine/PSC-833 as a P-gp substrate/inhibitor combination. The aim of the present study was to expand our assay system to mice using microdialysis with simultaneous sampling of blood and brain and to compare the result with a primary mouse brain endothelial cell monolayer (pMBMEC) assay. Brain penetration of quinidine was dose dependent in both anesthetized and awake mice after intraperitoneal drug administration. PSC-833 pretreatment caused a 2.5- to 3.4-fold increase in quinidine levels of brain dialysate samples in anesthetized or awake animals, after single or repeated administration of PSC-833. In pMBMEC, a 2.0- to 2.5-fold efflux ratio was observed in the transcellular transport of quinidine. The P-gp–mediated vectorial transport of quinidine was eliminated by PSC-833. These results indicate that quinidine with PSC-833 is a good probe substrate-reference inhibitor combination for testing drug-drug interactions with P-gp in the in vivo and in vitro mouse systems. With increasing number of humanized transgenic mice, a test system with mouse microdialysis experimentation becomes more important to predict drug-drug interactions in humans.

Quinidine as an ABCB1 Probe for Testing Drug Interactions at the Blood–Brain Barrier

2011 ◽  
Vol 16 (8) ◽  
pp. 886-894 ◽  
Author(s):  
István Sziráki ◽  
Franciska Erdő ◽  
Erzsébet Beéry ◽  
Petra Magdolna Molnár ◽  
Csilla Fazakas ◽  
...  
Keyword(s):  
Blood Brain Barrier ◽  
Cell System ◽  
Brain Barrier ◽  
In Vitro Assays ◽  
Specific Substrate ◽  
Psc 833 ◽  
Blood Brain ◽  
Substrate Inhibitor

This study provides evidence that quinidine can be used as a probe substrate for ABCB1 in multiple experimental systems both in vitro and in vivo relevant to the blood–brain barrier (BBB). The combination of quinidine and PSC-833 (valspodar) is an effective tool to assess investigational drugs for interactions on ABCB1. Effects of quinidine and substrate–inhibitor interactions were tested in a membrane assay and in monolayer assays. The authors compared quinidine and digoxin as ABCB1 probes in the in vitro assays and found that quinidine was more potent and at least as specific as digoxin in ATPase and monolayer efflux assays employing MDCKII-MDR1 and the rat brain microcapillary endothelial cell system. Brain exposure to quinidine was tested in dual-/triple-probe microdialysis experiments in rats by assessing levels of quinidine in blood and brain. Comparing quinidine levels in dialysate samples from valspodar-treated and control animals, it is evident that systemic/local administration of the inhibitor diminishes the pumping function of ABCB1 at the BBB, resulting in an increased brain penetration of quinidine. In sum, quinidine is a good probe to study ABCB1 function at the BBB. Moreover, quinidine/PSC-833 is an ABCB1-specific substrate/inhibitor combination applicable to many assay systems both in vitro and in vivo.

PET-CT imaging with [18F]-gefitinib to measure Abcb1a/1b (P-gp) and Abcg2 (Bcrp1) mediated drug–drug interactions at the murine blood–brain barrier

2015 ◽  
Vol 42 (11) ◽  
pp. 833-841 ◽  
Author(s):  
Maria L.H. Vlaming ◽  
Tilman Läppchen ◽  
Harm T. Jansen ◽  
Suzanne Kivits ◽  
Andy van Driel ◽  
...  
Keyword(s):  
Drug Interactions ◽  
Blood Brain Barrier ◽  
Ct Imaging ◽  
Brain Barrier ◽  
Blood Brain ◽  
Pet Ct

scholarly journals Drug interactions at the blood-brain barrier: Fact or fantasy?☆

2009 ◽  
Vol 123 (1) ◽  
pp. 80-104 ◽  
Author(s):  
Sara Eyal ◽  
Peng Hsiao ◽  
Jashvant D. Unadkat
Keyword(s):  
Drug Interactions ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Blood Brain

Rapid assessment of p-glycoprotein–drug interactions at the blood–brain barrier

2006 ◽  
Vol 358 (1) ◽  
pp. 51-58 ◽  
Author(s):  
Martin Bubik ◽  
Melanie Ott ◽  
Anne Mahringer ◽  
Gert Fricker
Keyword(s):  
Drug Interactions ◽  
Blood Brain Barrier ◽  
Rapid Assessment ◽  
Brain Barrier ◽  
P Glycoprotein ◽  
Blood Brain

scholarly journals Factors Governing P-Glycoprotein-Mediated Drug–Drug Interactions at the Blood–Brain Barrier Measured with Positron Emission Tomography

2015 ◽  
Vol 12 (9) ◽  
pp. 3214-3225 ◽  
Author(s):  
Thomas Wanek ◽  
Kerstin Römermann ◽  
Severin Mairinger ◽  
Johann Stanek ◽  
Michael Sauberer ◽  
...  
Keyword(s):  
Positron Emission Tomography ◽  
Drug Interactions ◽  
Blood Brain Barrier ◽  
Emission Tomography ◽  
Brain Barrier ◽  
P Glycoprotein ◽  
Blood Brain ◽  
Positron Emission

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

2009 ◽  
Vol 83 (18) ◽  
pp. 9398-9410 ◽  
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.

scholarly journals Brain endothelial tricellular junctions as novel sites for T cell diapedesis across the blood–brain barrier

2021 ◽  
Vol 134 (8) ◽  
Author(s):  
Mariana Castro Dias ◽  
Adolfo Odriozola Quesada ◽  
Sasha Soldati ◽  
Fabio Bösch ◽  
Isabelle Gruber ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Blood Brain Barrier ◽  
Immune Surveillance ◽  
Brain Barrier ◽  
Activated T Cells ◽  
Blood Brain ◽  
Primary Mouse ◽  
Underlying Mechanisms

ABSTRACT The migration of activated T cells across the blood–brain barrier (BBB) is a critical step in central nervous system (CNS) immune surveillance and inflammation. Whereas T cell diapedesis across the intact BBB seems to occur preferentially through the BBB cellular junctions, impaired BBB integrity during neuroinflammation is accompanied by increased transcellular T cell diapedesis. The underlying mechanisms directing T cells to paracellular versus transcellular sites of diapedesis across the BBB remain to be explored. By combining in vitro live-cell imaging of T cell migration across primary mouse brain microvascular endothelial cells (pMBMECs) under physiological flow with serial block-face scanning electron microscopy (SBF-SEM), we have identified BBB tricellular junctions as novel sites for T cell diapedesis across the BBB. Downregulated expression of tricellular junctional proteins or protein-based targeting of their interactions in pMBMEC monolayers correlated with enhanced transcellular T cell diapedesis, and abluminal presence of chemokines increased T cell diapedesis through tricellular junctions. Our observations assign an entirely novel role to BBB tricellular junctions in regulating T cell entry into the CNS. This article has an associated First Person interview with the first author of the paper.

scholarly journals BACE-1 is expressed in the blood–brain barrier endothelium and is upregulated in a murine model of Alzheimer’s disease

2015 ◽  
Vol 36 (7) ◽  
pp. 1281-1294 ◽  
Author(s):  
Kavi Devraj ◽  
Slobodan Poznanovic ◽  
Christoph Spahn ◽  
Gerhard Schwall ◽  
Patrick N Harter ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Endothelial Cells ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Blood Brain ◽  
Mouse Brain Endothelial Cells ◽  
Primary Mouse ◽  
Β Amyloid ◽  
Bace 1

Endothelial cells of the blood–brain barrier form a structural and functional barrier maintaining brain homeostasis via paracellular tight junctions and specific transporters such as P-glycoprotein. The blood–brain barrier is responsible for negligible bioavailability of many neuroprotective drugs. In Alzheimer’s disease, current treatment approaches include inhibitors of BACE-1 (β-site of amyloid precursor protein cleaving enzyme), a proteinase generating neurotoxic β-amyloid. It is known that BACE-1 is highly expressed in endosomes and membranes of neurons and glia. We now provide evidence that BACE-1 is expressed in blood–brain barrier endothelial cells of human, mouse, and bovine origin. We further show its predominant membrane localization by 3D- dSTORM super-resolution microscopy, and by biochemical fractionation that further shows an abluminal distribution of BACE-1 in brain microvessels. We confirm its functionality in processing APP in primary mouse brain endothelial cells. In an Alzheimer’s disease mouse model we show that BACE-1 is upregulated at the blood–brain barrier compared to healthy controls. We therefore suggest a critical role for BACE-1 at the blood–brain barrier in β-amyloid generation and in vascular aspects of Alzheimer’s disease, particularly in the development of cerebral amyloid angiopathy.

Sign in / Sign up

Export Citation Format

Share Document