20.500.12556/DiRROS-19380 Quantitative assessment of the apical and basolateral membrane expression of VEGFR2 and NRP2 in VEGF-A-stimulated cultured human umbilical vein endothelial cells Endothelial cells (ECs) form a precisely regulated polarized monolayer in capillary walls. Vascular endothelial growth factor-A (VEGF-A) induces endothelial hyperpermeability, and VEGF-A applied to the basolateral side, but not the apical side, has been shown to be a strong barrier disruptor in blood–retinal barrier ECs. We show here that VEGF-A presented to the basolateral side of human umbilical vein ECs (HUVECs) induces higher permeability than apical stimulation, which is similar to results obtained with bovine retinal ECs. We investigated with immunocytochemistry and confocal imaging the distribution of VEGF receptor-2 (VEGFR2) and neuropilin-2 (NRP2) in perinuclear apical and basolateral membrane domains. Orthogonal z-sections of cultured HUVECs were obtained, and the fluorescence intensity at the apical and basolateral membrane compartments was measured. We found that VEGFR2 and NRP2 are evenly distributed throughout perinuclear apical and basolateral membrane compartments in unstimulated HUVECs grown on Transwell inserts, whereas basolateral VEGF-A stimulation induces a shift toward basolateral VEGFR2 and NRP2 localization. When HUVECs were grown on coverslips, the distribution of VEGFR2 and NRP2 across the perinuclear apical and basolateral membrane domains was different. Our findings demonstrate that HUVECs dynamically regulate VEGFR2 and NRP2 localization on membrane microdomains, depending on growth conditions and the polarity of VEGF-A stimulation. endotelijske celice EGF celična biologija true false true Angleški jezik Ni določen Neznano 2024-07-17 03:55:51 2024-07-17 11:06:56 2024-07-18 03:53:59 0000-00-00 00:00:00 2022 0 0 Ostali avtorji: Shahan Darwesh, Jia Y. Zheng, Cornelis J.F. van Noorden, Reinier O. Schlingemann, and Ingeborg Klaassen; str. 557-569 št. 8 Vol. 70 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2022-08-01 576 0022-1554 10.1369/00221554221115767 121650947 https://doi.org/10.1369/00221554221115767 1 ee1fec70-43ce-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=26744 RAZ_Bosma_Esmeralda_K._2022.pdf RAZ_Bosma_Esmeralda_K._2022.pdf 1 EB48EBE47D389E3560F969BDD4E692A2 821764513624f9b1f27c1a9fb7cbbee5afed403dde73cc5c440baded9d1009e6 401278a8-440b-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=26829 Nacionalni inštitut za biologijo 0 0 0