Involvement of TRPV1 and TRPV4 Channels in Retinal Angiogenesis

Investigative Ophthalmology & Visual Science
Caitriona O'LearyTim M Curtis

Abstract

We investigate the contribution of TRPV1 and TRPV4 channels to retinal angiogenesis. 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. 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 angiog...Continue Reading

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Citations

Apr 10, 2020·Investigative Ophthalmology & Visual Science·Luka LapajneDavid Križaj
Apr 9, 2021·Biochemistry. Biokhimii︠a︡·Irina N GladkikhSergey A Kozlov
May 11, 2021·Microcirculation : the Official Journal of the Microcirculatory Society, Inc·Miao Chen, Xiucun Li
Sep 27, 2020·Experimental Eye Research·Brianna D GuarinoCharles K Thodeti

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