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α‐Adducin is crucial for endothelial Tight Junction Integrity
Author(s) -
Ponce Alexander García,
Radeva Mariya Yosifova,
Waschke Jens
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.686.3
Subject(s) - adherens junction , tight junction , microbiology and biotechnology , barrier function , cell junction , occludin , cytoskeleton , forskolin , endothelial stem cell , intracellular , actin cytoskeleton , biology , chemistry , cell , cell culture , cadherin , biochemistry , in vitro , genetics
Adducin (Add) is a membrane and cytoskeleton‐associated protein participating in the assembly of spectrin‐actin networks, which in turn regulate adherens junction (AJ) and tight junction (TJ) integrity. This process is important for endothelial barrier homeostasis. However, the mechanisms by which Add participates in endothelial barrier regulation are not well understood. Therefore, we generated immortalized Add knock‐out (KO) endothelial cell lines and investigated barrier dynamics. In absence of Add, transendothelial electrical resistance (TER) measurements showed impaired barrier function under basal conditions. Moreover, cAMP‐mediated barrier stabilization induced by treatment with Forskolin (F) and Rolipram (R) was compromised. Additionally, cells lacking Add showed higher intracellular cAMP concentration although the cAMP‐degrading enzyme phosphodiesterase 4D was overexpressed. Strikingly, ablation of Add led to a complete absence of TJ protein claudin‐5. Furthermore, in contrast to WT cells, Add KO cells displayed reduced membrane staining of other TJ molecule such as ZO‐1, both under resting conditions and after F/R treatment. However, the cellular distribution of this protein remained similar. In addition, localization of AJ protein VE‐cadherin at cell‐cell contacts was altered in Add KO cells, which was paralleled by significantly reduced protein levels. Taken together, in this study we show that Add is involved in endothelial barrier dynamics by regulating AJ and TJ integrity as well as cAMP‐mediated‐signalling This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .