title: Vascular endothelial growth factor-C, a potential paracrine regulator of glomerular permeability, increases glomerular endothelial cell monolayer integrity and intracellular calcium.
abstract: We have previously reported expression of vascular endothelial growth factor (VEGF)-A and -C in glomerular podocytes and actions of VEGF-A on glomerular endothelial cells (GEnC) that express VEGF receptor-2 (VEGFR-2). Here we define VEGFR-3 expression in GEnC and investigate the effects of the ligand VEGF-C. Renal cortex and cultured GEnC were examined by microscopy, and both cell and glomerular lysates were assessed by Western blotting. VEGF-C effects on trans-endothelial electrical resistance and albumin flux across GEnC monolayers were measured. The effects of VEGF-C156S, a VEGFR-3-specific agonist, and VEGF-A were also studied. VEGF-C effects on intracellular calcium ([Ca2+]i) were measured using a fluorescence technique, receptor phosphorylation was examined by immunoprecipitation assays, and phosphorylation of myosin light chain-2 and VE-cadherin was assessed by blotting with phospho-specific antibodies. GEnC expressed VEGFR-3 in tissue sections and culture, and VEGF-C increased trans-endothelial electrical resistance in a dose-dependent manner with a maximal effect at 120 minutes of 6.8 Omega whereas VEGF-C156S had no effect. VEGF-C reduced labeled albumin flux by 32.8%. VEGF-C and VEGF-A increased [Ca2+]i by 15% and 39%, respectively. VEGF-C phosphorylated VEGFR-2 but not VEGFR-3, myosin light chain-2, or VE-cadherin. VEGF-C increased GEnC monolayer integrity and increased [Ca2+]i, which may be related to VEGF-C-S particular receptor binding and phosphorylation induction characteristics. These observations suggest that podocytes direct GEnC behavior through both VEGF-C and VEGF-A.
authors.affiliation: Academic Renal Unit, Southmead Hospital, Bristol, UK.
authors.forename: Rebecca R
authors.initials: RR
authors.lastname: Foster
authors.forename: Sadie C
authors.initials: SC
authors.lastname: Slater
authors.forename: Jaqualine
authors.initials: J
authors.lastname: Seckley
authors.forename: Dontscho
authors.initials: D
authors.lastname: Kerjaschki
authors.forename: David O
authors.initials: DO
authors.lastname: Bates
authors.forename: Peter W
authors.initials: PW
authors.lastname: Mathieson
authors.forename: Simon C
authors.initials: SC
authors.lastname: Satchell
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D024722:Cardiac Myosins / Q000378:metabolism; D002460:Cell Line; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D050505:Mutant Proteins / Q000494:pharmacology; D018994:Myosin Light Chains / Q000378:metabolism; D019899:Paracrine Communication / Q000187:drug effects*; D010539:Permeability / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D042582:Vascular Endothelial Growth Factor C / Q000494:pharmacology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism / Q000648:ultrastructure; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D002118:Calcium; D024722:Cardiac Myosins; D016650:Fluorescein-5-isothiocyanate; D050505:Mutant Proteins; D018994:Myosin Light Chains; D042461:Vascular Endothelial Growth Factor A; D042582:Vascular Endothelial Growth Factor C; D040301:Vascular Endothelial Growth Factor Receptor-2; D040321:Vascular Endothelial Growth Factor Receptor-3; D000099152:Cadherin 5; C107184:myosin light chain 2
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