Abstract
This study was performed to determine whether the α subunit of ATP synthase (α-ATP synthase) on brain microvascular endothelial cells (BMECs) serves as the functional target for endothelial monocyte-activating polypeptide-II (EMAP-II)-induced increase in blood-tumor barrier (BTB) permeability. Using a rat C6 glioma model, we found that low-dose (80 ng/kg) EMAP-II significantly decreased the mRNA and protein expression levels of tight junction (TJ)-related proteins claudin-5, occludin, and ZO-1 on BMECs. Meantime, radioimmunity and Western blot assay showed a significant decrease in the expression levels of cAMP and catalytic subunit of protein kinase A (PKAcs) of tumor tissues. Also, pretreatment with specific α-ATP synthase antibody significantly blocked the effects of EMAP-II on TJ-related proteins, cAMP, and PKAcs. In addition, double immunofluorescence assay identified that EMAP-II was co-localized with α-ATP synthase on BMECs. This in vivo study demonstrated that α subunit of ATP synthase on BMECs serves as the functional target for EMAP-II selective opening of the BTB, and that cAMP/PKA signaling transduction pathway might be involved in the modulating process.
Copyright © 2010 Elsevier B.V. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Brain / blood supply
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Brain / metabolism
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Capillary Permeability / drug effects
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Capillary Permeability / physiology*
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Cell Line, Tumor
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Claudin-5
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Cyclic AMP / metabolism
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Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / metabolism
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Cytokines / pharmacology*
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Disease Models, Animal
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Endothelial Cells / drug effects
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Endothelial Cells / metabolism
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Evans Blue / metabolism
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Glioma / blood supply
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Glioma / metabolism*
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Male
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Membrane Proteins / metabolism
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Mitochondrial Proton-Translocating ATPases / antagonists & inhibitors
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Mitochondrial Proton-Translocating ATPases / physiology*
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Neoplasm Proteins / pharmacology*
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Occludin
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Phosphoproteins / metabolism
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Protein Subunits / antagonists & inhibitors
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Protein Subunits / physiology
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Proton-Translocating ATPases / antagonists & inhibitors
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Proton-Translocating ATPases / drug effects
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Proton-Translocating ATPases / physiology*
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RNA-Binding Proteins / pharmacology*
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Rats
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Rats, Wistar
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Zonula Occludens-1 Protein
Substances
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Claudin-5
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Cldn5 protein, rat
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Cytokines
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Membrane Proteins
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Neoplasm Proteins
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Occludin
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Ocln protein, rat
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Phosphoproteins
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Protein Subunits
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RNA-Binding Proteins
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Tjp1 protein, rat
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Zonula Occludens-1 Protein
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small inducible cytokine subfamily E, member 1
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Evans Blue
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Cyclic AMP
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Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
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Mitochondrial Proton-Translocating ATPases
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Proton-Translocating ATPases