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Papers In Press, published online ahead of print February 14, 2002
Department of Anethesia, Brigham and Women's Hospital, Boston, MA 02115
Corresponding Author: colgan{at}zeus.bwh.harvard.edu
Endothelial barrier function is altered by the release of soluble polymorphonuclear leukocyte (PMN)-derived mediators during inflammatory states. However, endogenous pathways to describe such changes are only recently appreciated. Using an in vitro endothelial paracellular permeability model, cell-free supernatants from FMLP-stimulated PMN were observed to significantly alter endothelial permeability. Biophysical and biochemical analysis of PMN supernatants identified PMN-derived glutamate in modulating endothelial permeability. Further, novel expression of metabotropic glutamate receptor 1 (mGluR1), mGluR4 and mGluR5 by human brain and dermal microvascular endothelial cells was demonstrated by RT-PCR, in situ hybridization, immunofluorescence and western blot analysis. Treatment of human brain endothelia with glutamate or selective mGluR group I or III agonists resulted in a time-dependent loss of phosphorylated vasodilator-stimulated phosphoprotein (VASP), and significantly increased endothelial permeability. Glutamate-induced decreases in brain endothelial barrier function and phosphorylated VASP were significantly attenuated by pre-treatment of human brain endothelia with slective mGluR antagonists. These observations were extended to an in vivo hypoxic mouse model in which pre-treatment with mGluR antagonists significantly decreased FITC-dextran flux across the blood-brain barrier. We conclude that activated human PMN release glutamate and that endothelial expression of group I or III mGluR function to decrease human brain endothelial VASP phosphorylation and barrier function. These results identify a novel pathway by which PMN-derived glutamate may regulate human endothelial barrier function.
J. Biol. Chem, 10.1074/jbc.M110557200
Submitted on November 2, 2001
Revised on February 13, 2002
Accepted on February 14, 2002
Neutrophil-derived glutamate regulates vascular endothelial barrier function
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