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您的检索式：作者名="Ariel M.Wilson"
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| 1 | AB018.Ocular hypertension promotes early mitochondrial fragmentation in retinal endothelial cells in a mouse model of glaucoma显示文摘Background:Retinal endothelial cells are very active and contribute to the integrity of the neurovascular unit.Vascular dysfunction has been proposed to contribute to the pathogenesis of glaucoma.Here,we evaluated the hypothesis that ocular hypertension triggers mitochondrial alterations in endothelial cells impairing the integrity of the blood retinal barrier(BRB).Methods:Ocular hypertension was induced by injection of magnetic microbeads into the anterior chamber of EndoMito-EGFP mice,a strain expressing green fluorescent protein selectively in the mitochondria of endothelial cells.Capillary density,mitochondrial volume,and the number of mitochondrial components were quantified in 3D-reconstructed images from whole-mounted retinas using Imaris software.Dynamin-related protein(DRP-1),mitofusin-2(MFN-2)and optic atrophy-1(OPA-1)expression were assessed by western blot analysis of enriched endothelial cells.Mitochondrial structure was evaluated by transmission electron microscopy(TEM)and oxygen consumption rate was monitored by Seahorse analysis.The integrity of the BRB was evaluated by quantifying Evans blue leakage.Results:Our data demonstrate that two and three weeks after ocular hypertension induction,the total mitochondria volume in endothelial cells decreased from 0.140±0.002µm3 from non-injured retinas to 0.108±0.005 and 0.093±0.007µm3,respectively in glaucomatous eyes(mean±S.E.M,ANOVA,P<0.001;N=6/group).Frequency distribution showed a substantial increase of smaller mitochondria complexes(<0.5µm3)in endothelial cells from glaucomatous retinas.Significant upregulation of DRP-1 was found in vessels isolated from glaucomatous retinas compared to the intact retinas,while MFN-2 and OPA-1 expression was not affected.Structural alteration in endothelial cell mitochondria was confirmed by TEM,which were accompanied by a 1.93-fold reduction in the oxygen consumption rate as well as 2.6-fold increase in vasculature leakage in glaucomatous retinas(n=3-6/group).In addition,this model did not trigger changes in the density of the vascular network,suggesting that mitochondrial fragmentation was not due to endothelial cell loss.Conclusions:This study shows that ocular hypertension leads to early alterations in the dynamic of endothelial cell mitochondria,contributing to vascular dysfunction in glaucoma. | Jorge L.Cueva Vargas Yoko Ito Khalil Miloudi Ariel M.Wilson Christine Vande Velde Przemyslaw Sapieha Adriana Di Polo | 2018 | Annals of Eye Science2018，，1： | 0 |
| 2 | AB034. In vivo laser-mediated retinal ganglion cell optoporation using Kv1.1 conjugated gold nanoparticles显示文摘Background:There is a current void in efficient,cell-specific,retinal drug delivery systems,thus developing a safe,effective,selective drug delivery system would open novel therapeutic avenues.We previously demonstrated that femtosecond(fs)laser irradiation can transfect DNA plasmids into cultured cells in the presence of gold nanoparticles(AuNPs).These AuNPs locally amplify laser energy at a submicron range creating transient pores allowing exogenous genetic material or cell impermeable dyes to enter the cell.Here,we sought out to selectively optoporate retinal cells in vivo with functionalized AuNPs and a 800 nm femtosecond(fs)laser.Methods:The cell-surface Kv1.1 voltage-gated channel was chosen to selectively target retinal ganglion cells(RGC)in the rat retina.Citrate-capped spherical 100 nm AuNPs functionalized with orthopyridyl-disulfide-poly(ethylene glycol)(5 kDa)-N-hydroxysuccinimide(OPSS-PEG-NHS)conjugated to a Kv1.1 monoclonal antibody were injected intravitreally in Sprague Dawley rats 3 hours prior to irradiation,concomitantly to a FITC-dextran dye to detect optoporation.The eyes of anesthetized rats were placed in the beam path of a laser system consisting of an 800 nm,100 fs laser and a Heidelberg Spectralis HRA ophthalmoscope for fundus visualization.The rat retina was irradiated at powers ranging from 20-750 mW,the eyes fixed in 4%paraformaldehyde,dissected,rinsed,mounted and imaged by confocal microscopy.Results:Our novel laser system coupled to a Heidelberg ophthalmoscope allowed for a clear visualisation of the rat ocular fundus.A timecourse of AuNP intravitreal injections revealed that optimal nanoparticle dispersion on the retinal surface occurred at 3 hours post injection.Following Kv1.1-AuNP and FITC-dextran intravitreal injection and incubation,irradiation at 120-750 mW resulted in FITC uptake by retinal cells.Conclusions:Since living biological tissues absorb energy very weakly at 800 nm,this non-invasive tool may provide a safe,cost effective clinically relevant approach to selectively target retinal cells and limit complications associated with surgical interventions,and potential biological hazards associated with viral-based gene therapy. | Ariel M.Wilson Javier Mazzaferri Éric Bergeron Sergiy Patskovsky Santiago Costantino Michel Meunier Przemyslaw Sapieha | 2018 | Annals of Eye Science2018，，1： | 0 |
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