dc.date.accessioned | 2020-07-10T17:57:21Z | |
dc.date.available | 2020-07-10T17:57:21Z | |
dc.date.created | 2019-11-24T17:26:21Z | |
dc.date.issued | 2019 | |
dc.identifier.citation | Mousavi, Seyed Ali Skjeldal, Frode Miltzow Fønhus, Marita Sporstøl Haugen, Linda Hofstad Eskild, Winnie Berg, Trond Bakke, Oddmund . Receptor-mediated endocytosis of VEGF-A in rat liver sinusoidal endothelial cells. BioMed Research International. 2019, 2019:5496197, 1-12 | |
dc.identifier.uri | http://hdl.handle.net/10852/77747 | |
dc.description.abstract | Background and Aims. Vascular endothelial growth factor (VEGF) receptors (VEGFR1 and VEGFR2) bind VEGF-A with high affinity. This study sought to determine the relative contributions of these two receptors to receptor-mediated endocytosis of VEGF-A and to clarify their endocytic itineraries in rat liver sinusoidal endothelial cells (LSECs). Methods. Isolated LSECs and radiolabeled VEGF-A were used to examine surface binding and receptor-mediated endocytosis. Quantitative real time RT-PCR (Q-RT-PCR) and Western blotting were applied to demonstrate receptor expression. Results. Q-RT-PCR analysis showed that VEGFR1 and VEGFR2 mRNA were expressed in LSECs. Ligand saturation analysis at 4°C indicated two different classes of [125I]-VEGFA binding sites on LSECs with apparent dissociation constants of 8 and 210 pM. At 37°C, LSECs efficiently took up and degraded [125I]-VEGF-A for at least 2 hours. Uptake of [125I]-VEGF-A by LSECs was blocked by dynasore that inhibits dynamin-dependent internalization, whereas inhibition of cysteine proteases by leupeptin inhibited degradation without affecting the uptake of [125I]-VEGF-A, suggesting that it is degraded following transport to lysosomes. Incubation of LSECs in the continued presence of a saturating concentration of unlabeled VEGF-A at 37°C was associated with a loss of as much as 75% of the total VEGFR2 within 30 min as shown by Western blot analysis, whereas there was no appreciable decrease in protein levels for VEGFR1 after 120 min incubation, suggesting that VEGF-A stimulation downregulates VEGFR2, but not VEGFR1, in LSECs. This possibility was supported by the observation that a hexapeptide that specifically blocks VEGF-A binding to VEGFR1 caused a marked reduction in the uptake of [125I]-VEGF-A, whereas a control peptide had no effect. Finally, live cell imaging studies using a fluorescently labeled anti-VEGFR2 antibody showed that VEGFR2 was transported via early and late endosomes to reach endolysosomes where degradation of the VEGFR2 takes place. Conclusion. Our studies suggest that, subsequent to VEGF-A binding and internalization, the unoccupied VEGFR1 may recycle to the cell surface allowing its reutilization, whereas the majority of the internalized VEGFR2 is targeted for degradation. | |
dc.language | EN | |
dc.rights | Attribution 4.0 International | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
dc.title | Receptor-mediated endocytosis of VEGF-A in rat liver sinusoidal endothelial cells | |
dc.type | Journal article | |
dc.creator.author | Mousavi, Seyed Ali | |
dc.creator.author | Skjeldal, Frode Miltzow | |
dc.creator.author | Fønhus, Marita Sporstøl | |
dc.creator.author | Haugen, Linda Hofstad | |
dc.creator.author | Eskild, Winnie | |
dc.creator.author | Berg, Trond | |
dc.creator.author | Bakke, Oddmund | |
cristin.unitcode | 185,15,29,30 | |
cristin.unitname | Seksjon for fysiologi og cellebiologi | |
cristin.ispublished | true | |
cristin.fulltext | original | |
cristin.qualitycode | 1 | |
dc.identifier.cristin | 1751502 | |
dc.identifier.bibliographiccitation | info:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=BioMed Research International&rft.volume=2019:5496197&rft.spage=1&rft.date=2019 | |
dc.identifier.jtitle | BioMed Research International | |
dc.identifier.volume | 2019 | |
dc.identifier.doi | https://doi.org/10.1155/2019/5496197 | |
dc.identifier.urn | URN:NBN:no-80873 | |
dc.type.document | Tidsskriftartikkel | |
dc.type.peerreviewed | Peer reviewed | |
dc.source.issn | 2314-6133 | |
dc.identifier.fulltext | Fulltext https://www.duo.uio.no/bitstream/handle/10852/77747/2/Receptor-Mediated%2BEndocytosis-5496197.pdf | |
dc.type.version | PublishedVersion | |
cristin.articleid | 5496197 | |
dc.relation.project | NFR/230779 | |