Inhibition of Glycogen Synthase Kinase 3β Promotes Tight Junction Stability in Brain Endothelial Cells by Half-Life Extension of Occludin and Claudin-5

Ramirez, Servio H.; Fan, Shongshan; Dykstra, Holly; Rom, Slava; Mercer, Aaron; Reichenbach, Nancy L.; Gofman, Larisa; Persidsky, Yuri
February 2013
PLoS ONE;Feb2013, Vol. 8 Issue 2, p1
Academic Journal
Neuroinflammatory conditions often involve dysfunction of the Blood-Brain Barrier (BBB). Therefore, identifying molecular targets that can maintain barrier fidelity is of clinical importance. We have previously reported on the anti-inflammatory effects that glycogen synthase kinase 3β (GSK3β) inhibition has on primary human brain endothelial cells. Here we show that GSK3β inhibitors also promote barrier tightness by affecting tight junction (TJ) protein stability. Transendothelial electrical resistance (TEER) was used to evaluate barrier integrity with both pharmacological inhibitors and mutants of GSK3β. Inhibition of GSK3β produced a gradual and sustained increase in TEER (as much as 22% over baseline). Analysis of subcellular membrane fractions revealed an increase in the amount of essential tight junction proteins, occludin and claudin-5, but not claudin-3. This phenomenon was attributed to a decrease in TJ protein turnover and not transcriptional regulation. Using a novel cell-based assay, inactivation of GSK3β significantly increased the half-life of occludin and claudin-5 by 32% and 43%, respectively. A correlation was also established between the enhanced association of β-catenin with ZO-1 as a function of GSK3β inhibition. Collectively, our findings suggest the possibility of using GSK3β inhibitors as a means to extend the half-life of key tight junction proteins to promote re-sealing of the BBB during neuroinflammation.


Related Articles

  • Alterations of Blood-Brain Barrier and Associated Factors in Acute Liver Failure. Wei Cui; Cui-Ming Sun; Pei Liu // Gastroenterology Research & Practice;2013, p1 

    Brain edema in acute liver failure (ALF) remains lethal. Cytotoxic mechanisms associated with brain edema have been well recognized, but the role of vasogenic mechanisms of brain edema has not been explored. Intact tight junctions (TJs) between brain capillary endothelial cells are critical for...

  • Molecular and structural transmembrane determinants critical for embedding claudin-5 into tight junctions reveal a distinct four-helix bundle arrangement. Rossa, Jan; Protze, Jonas; Kern, Christian; Piontek, Anna; Günzel, Dorothee; Krause, Gerd; Piontek, Jörg // Biochemical Journal;11/15/2014, Vol. 464 Issue 1, p49 

    The mechanism of TJ (tight junction) assembly and the structure of TJ strand-forming Cldns (claudins) are unclear. To identify determinants of assembly of blood-brain barrier-related Cldn3 and Cldn5, chimaeric mutants were analysed by cellular reconstitution of TJ strands and live-cell imaging....

  • Proteomic Analysis of Proteins Surrounding Occludin and Claudin-4 Reveals Their Proximity to Signaling and Trafficking Networks. Fredriksson, Karin; Van Itallie, Christina M.; Aponte, Angel; Gucek, Marjan; Tietgens, Amber J.; Anderson, James M. // PLoS ONE;Mar2015, Vol. 10 Issue 3, p1 

    Tight junctions are complex membrane structures that regulate paracellular movement of material across epithelia and play a role in cell polarity, signaling and cytoskeletal organization. In order to expand knowledge of the tight junction proteome, we used biotin ligase (BioID) fused to occludin...

  • Cooling Treatment Transiently Increases the Permeability of Brain Capillary Endothelial Cells Through Translocation of Claudin-5. Inamura, Akinori; Adachi, Yasuhiro; Inoue, Takao; He, Yeting; Tokuda, Nobuko; Nawata, Takashi; Shirao, Satoshi; Nomura, Sadahiro; Fujii, Masami; Ikeda, Eiji; Owada, Yuji; Suzuki, Michiyasu // Neurochemical Research;Aug2013, Vol. 38 Issue 8, p1641 

    The blood-brain-barrier (BBB) is formed by different cell types, of which brain microvascular endothelial cells are major structural constituents. The goal of this study was to examine the effects of cooling on the permeability of the BBB with reference to tight junction formation of brain...

  • Small molecules enable highly efficient neuronal conversion of human fibroblasts. Ladewig, Julia; Mertens, Jerome; Kesavan, Jaideep; Doerr, Jonas; Poppe, Daniel; Glaue, Finnja; Herms, Stefan; Wernet, Peter; Kögler, Gesine; Müller, Franz-Josef; Koch, Philipp; Brüstle, Oliver // Nature Methods;Jun2012, Vol. 9 Issue 6, p575 

    Forced expression of proneural transcription factors has been shown to direct neuronal conversion of fibroblasts. Because neurons are postmitotic, conversion efficiencies are an important parameter for this process. We present a minimalist approach combining two-factor neuronal programming with...

  • Hypercholesterolaemic Serum Increases the Permeability of Endothelial Cells through Zonula Occludens-1 with Phosphatidylinositol 3-Kinase Signaling Pathway. Chang Bian; Geng Xu; Jianan Wang; Ji Ma; MeiXiang Xiang; Peng Chen // Journal of Biomedicine & Biotechnology;2009 Regular Issue, p1 

    Purpose. Hypercholesterolemia and tight junctions play important roles in atherosclerosis. But the relationship between these two factors is unclear. In the present study, we investigated whether hypercholesterolemic serum could change the permeability of endothelial cells through altering...

  • Claudin-2 Regulates Colorectal Inflammation via Myosin Light Chain Kinase-Dependent Signaling. Nishida, Masayuki; Yoshida, Masaru; Nishiumi, Shin; Furuse, Mikio; Azuma, Takeshi // Digestive Diseases & Sciences;Jun2013, Vol. 58 Issue 6, p1546 

    Background: Claudins have been demonstrated to be associated with inflammatory bowel disease (IBD), but the specific role of claudin-2 in colorectal inflammation remains undefined. Aims: We aimed to determine the role of claudin-2 in TNFα-induced colorectal inflammation. Methods: We used...

  • C-type natriuretic peptide modulates permeability of the blood-brain barrier. Bohara, Manoj; Kambe, Yuki; Nagayama, Tetsuya; Tokimura, Hiroshi; Arita, Kazunori; Miyata, Atsuro // Journal of Cerebral Blood Flow & Metabolism;Apr2014, Vol. 34 Issue 4, p589 

    C-type natriuretic peptide (CNP) is abundant in brain and is reported to exert autocrine function in vascular cells, but its effect on blood-brain barrier (BBB) permeability has not been clarified yet. Here, we examined this effect. Transendothelial electrical resistance (TEER) of in vitro BBB...

  • Interleukin-34 Restores Blood–Brain Barrier Integrity by Upregulating Tight Junction Proteins in Endothelial Cells. Jin, Shijie; Sonobe, Yoshifumi; Kawanokuchi, Jun; Horiuchi, Hiroshi; Cheng, Yi; Wang, Yue; Mizuno, Tetsuya; Takeuchi, Hideyuki; Suzumura, Akio // PLoS ONE;Dec2014, Vol. 9 Issue 12, p1 

    Interleukin-34 (IL-34) is a newly discovered cytokine as an additional ligand for colony stimulating factor-1 receptor (CSF1R), and its functions are expected to overlap with colony stimulating factor-1/macrophage-colony stimulating factor. We have previously shown that the IL-34 is primarily...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics