Introduction In diabetic nephropathy, increased VEGFA expression is associated with proteinuria [1], however blocking VEGFA therapeutically also blocks its beneficial effects [2]. Previously we have shown that, in contrast to VEGFA, VEGFC decreases protein passage in human glomerular endothelial cells (GEnC) and tyrosine phosphorylates the same receptor as VEGFA, VEGFR2 [3]. Our aims were to determine how VEGFC induces differential signaling effects to VEGFA in GEnC in culture and in a transgenic mouse model, and the impacts of this on GEnC health. Methods A podocyte-specific, inducible VEGFC overexpressing mouse (podVEGFC) was developed by crossing PodrtTA mice with TetO-VEGFC mice. Mice were either terminally anaesthetised (Dormitor 1.6mg/kg, Narkitan 100mg/kg, H2O (1:1:1) i.m.) or killed by schedule 1. Glomeruli were isolated from podVEGFC mice using dynabeads as previously [4], RNA extracted, reverse transcribed and QPCR performed for VEGFC, VEGFR2, VEGFA and GAPDH. PodVEGFC kidneys were wax embedded, sectioned and stained with Periodic Acid Schiff, or snap frozen and sectioned. Kidneys were also fixed using cardiac perfusion with gluteradehyde, processed and imaged by electron microscopy (EM). VEGFR2/R3 heterodimerisation was analysed in fresh frozen kidney using a proximity ligation assay (PLA). Cultured GEnC were stimulated with vehicle, 1nM VEGFA or 10nM mature VEGFC for varying times and either stained with trypan blue and counted or protein/RNA extracted. Protein was Western blotted and probed for specific phospho-/pan VEGFR2 or phospho-/total Akt antibodies. QPCR was performed for VEGFR2 and GAPDH. Results VEGFC treatment did not phosphorylate VEGFR2 at tyrosine residues 1175, 1214 or 951 in GEnC (min. n=7). QPCR demonstrated that glomerular VEGFR2 (0.39±0.155 fold decrease) and VEGFA (0.62± 0.1 fold decrease, p<0.05, n=3) expression was decreased in podVEGFC mice and VEGFR2 was also decreased in GEnC stimulated with rhVEGFC (0.19±0.01 fold decrease compared to vehicle). A PLA demonstrated increased VEGFR2/3 heterodimerisation in podVEGFC mouse glomeruli (p<0.01, n=3). VEGFC promoted decreased trypan blue positive GEnC over 24h (p<0.05, n=5), increased Akt phosphorylation (n=3) and there were no morphological changes by light (min. n= 5) or EM (n=4) in podVEGFC glomeruli. Conclusions Although VEGFA and VEGFC both induce VEGFR2 phosphorylation in GEnC, VEGFC affects VEGFR2 and VEGFA expression and activates VEGFR2/R3 heterodimerisation in glomeruli ex vivo. Both VEGFC and VEGFA promote GEnC cell survival, and glomerular VEGFC overexpression has no detrimental effects on glomerular filtration barrier structure or function. Since VEGFC can reduce protein passage through GEnC monolayers, these results indicate potential therapeutic benefits of glomerular VEGFC on proteinuric kidney disease.