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Focal Adhesion Kinase Mediates Nucleotide P2Y 2 Receptor‐Dependent Increased Endothelial Permeability
Author(s) -
Wang Jianjie,
Harvey Joseph,
Tiwari Aparna,
Erb Laurie,
Huxley Virginia,
Weisman Gary,
Richard Garad
Publication year - 2015
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.29.1_supplement.791.9
Subject(s) - adherens junction , focal adhesion , tyrosine kinase , ve cadherin , tyrosine phosphorylation , receptor , chemistry , p2y receptor , extracellular , proto oncogene tyrosine protein kinase src , phosphorylation , microbiology and biotechnology , cadherin , biology , agonist , biochemistry , cell
Extracellular nucleotides play important roles in inflammation. We previously found that nucleotide P2Y 2 receptor activation induced transient 5‐fold increase in venular permeability to albumin ( P s ) from baseline in wild type (WT) mice, but not in P2Y 2 R knockout (P2Y 2 R KO) mice. The objective of this study is to investigate the role of focal adhesion kinase (FAK) in P2Y 2 R signaling‐dependent modulation of endothelial permeability. Primary cultured microvascular endothelial cells (MEC) of skeletal muscles, derived from either WT or P2Y 2 R KO mice, were used in this study. Immunoblot analysis revealed increase in FAK activity of 1.48±0.11 (n=3) and 1.58±0.08 (n=3) at 10 min and 30 min UTP (an agonist of P2Y 2 R; 10 ‐5 M), respectively, in WT MEC. In contrast, FAK activity didn't change in P2Y 2 R KO MEC. Activity of VE‐cadherin, a key molecule of the adherens junction, was assessed via measuring tyrosine phosphorylation of VE‐cadherin at tyrosine 658 (pY658) in HUVEC. UTP (10 ‐5 M) induced 1.16±0.03 (n=4) and 1.19±0.07 (n=4) increase in pY658 level of VE‐cadherin at 5 and 15 min, respectively, compared to the control group. The findings support the hypothesis that FAK activation is associated with P2Y 2 R‐induced increased microvascular permeability. (Supported by Missouri State University Research Funding).