Involvement of TRPV1 and TRPV4 Channels in Retinal Angiogenesis
| Autor: | Thomas Friedel, J. Graham McGeown, Sadaf Ashraf, Jennifer McNaughten, José A Fernández, Patrizia Cincolà, Mary K. McGahon, Alan W. Stitt, Peter Barabas, Tim M. Curtis, Caitriona O'Leary |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Vascular Endothelial Growth Factor A
0301 basic medicine TRPV4 Patch-Clamp Techniques Pyridines Angiogenesis Blotting Western TRPV Cation Channels Retinal Neovascularization Real-Time Polymerase Chain Reaction Mice 03 medical and health sciences chemistry.chemical_compound Immunolabeling 0302 clinical medicine Cell Movement In vivo medicine Animals Calcium Signaling RNA Messenger Sulfones Patch clamp Cells Cultured Cell Proliferation Sulfonamides Retina Endothelial Cells Retinal Vessels Retinal Cell migration General Medicine Cell biology Mice Inbred C57BL Oxygen 030104 developmental biology medicine.anatomical_structure Animals Newborn chemistry Calcium lipids (amino acids peptides and proteins) 030217 neurology & neurosurgery |
| Zdroj: | O'Leary, C, McGahon, M K, Ashraf, S, McNaughten, J, Friedel, T, Cincolà, P, Barabas, P, Fernandez, J A, Stitt, A W, McGeown, J G & Curtis, T M 2019, ' Involvement of TRPV1 and TRPV4 Channels in Retinal Angiogenesis ', Investigative ophthalmology & visual science, vol. 60, no. 10, pp. 3297-3309 . https://doi.org/10.1167/iovs.18-26344 |
| DOI: | 10.1167/iovs.18-26344 |
| Popis: | Purpose: We investigate the contribution of TRPV1 and TRPV4 channels to retinal angiogenesis.Methods: Primary retinal microvascular endothelial cells (RMECs) were used for RT-PCR, Western blotting, immunolabeling, Ca2+ signaling, and whole-cell patch-clamp studies while localization of TRPV1 also was assessed in retinal endothelial cells using whole mount preparations. The effects of pharmacologic blockers of TRPV1 and TRPV4 on retinal angiogenic activity was evaluated in vitro using sprout formation, cell migration, proliferation, and tubulogenesis assays, and in vivo using the mouse model of oxygen-induced retinopathy (OIR). Heteromultimerization of TRPV1 and TRPV4 channels in RMECs was assessed using proximity ligation assays (PLA) and electrophysiologic recording.Results: TRPV1 mRNA and protein expression were identified in RMECs. TRPV1 labelling was found to be mainly localized to the cytoplasm with some areas of staining colocalizing with the plasma membrane. Staining patterns for TRPV1 were broadly similar in endothelial cells of intact vessels within retinal flat mounts. Functional expression of TRPV1 and TRPV4 in RMECs was confirmed by patch-clamp recording. Pharmacologic inhibition of TRPV1 or TRPV4 channels suppressed in vitro retinal angiogenesis through a mechanism involving the modulation of tubulogenesis. Blockade of these channels had no effect on VEGF-stimulated angiogenesis or Ca2+ signals in vitro. PLA and patch-clamp studies revealed that TRPV1 and TRPV4 form functional heteromeric channel complexes in RMECs. Inhibition of either channel reduced retinal neovascularization and promoted physiologic revascularization of the ischemic retina in the OIR mouse model.Conclusions: TRPV1 and TRPV4 channels represent promising targets for therapeutic intervention in vasoproliferative diseases of the retina. |
| Databáze: | OpenAIRE |
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