The gene expression of NMDAR was assessed by RT-qPCR, and human neuroblastoma cells were used as a positive control. NMDAR1 expression was assessed in the human RPE (ARPE-19) cell line at both the gene and protein levels. Homocysteine Activates HIF-1α and VEGF in RPE Cells Moreover, to confirm the effect of Hcy treatment on the proliferation of choroidal endothelial cells, we examined the expression of the Ki67 proliferation factor in cultured mouse choroidal endothelial cells (MCEC), and results showed a marked increase in Ki67 expression (green) in MCEC by Hcy treatment ( Figure 1f). Retinal flat-mounts of mice that were exposed to laser induction with/without the intravitreal injection of Hcy were stained with isolectin-B4 to examine the effect of Hcy on the size and extent of laser-induced CNV this showed that Hcy significantly increased the extent of laser-induced CNV in wild-type mice ( Figure 1d,e). The results showed that Hcy-injected wild-type mice highly expressed isolectin-B4 and showed the development of blood vessels extending from the area of the choroid to the inner retina (white arrows, Figure 1c) as compared to the control wild-type mice, which revealed a normal retinal vascular pattern. Retinal section/flat-mounts were evaluated by immunofluorescence staining for vascular marker, isolectin-B4 (red). To study the effect of Hcy on CNV induction, we examined the CNV in the retinal frozen section and retinal flat-mounts of wild-type mice (C57-BL6) injected intravitreally with/without Hcy. Our findings underscore the role of RPE NMDAR in Hcy-induced AMD features thus, NMDAR inhibition could serve as a promising therapeutic target for AMD. Knocking down RPE NMDAR improved retinal structure and CNV. Hcy-injected WT mice showed disrupted retina and CNV. Hcy significantly increased HIF-1α, VEGF, and NMDAR in RPE cells, and Ki67 in MCEC. A neovascular AMD patient’s serum showed a significant increase in Hcy and a decrease in CBS. Isolectin-B4, Ki67, HIF-1α, VEGF, NMDAR1, and albumin were assessed by immunofluorescence (IF), Western blot (WB), Optical coherence tomography (OCT), and fluorescein angiography (FA) to evaluate retinal structure, fluorescein leakage, and choroidal neovascularization (CNV).

The involvement of NMDAR in Hcy-induced AMD features was evaluated (1) in vitro using ARPE-19 cells, primary RPE isolated from HHcy mice (CBS), and mouse choroidal endothelial cells (MCEC) (2) in vivo using wild-type mice and mice deficient in RPE NMDAR ( NMDAR R −/−) with/without Hcy injection. Serum Hcy and cystathionine-β-synthase (CBS) were assessed by ELISA. This study suggests that N-Methyl- d-aspartate receptor (NMDAR) activation in the retinal pigment epithelium (RPE) is a mechanism for HHcy-induced AMD.

We previously reported that HHcy induces AMD-like features. Elevated homocysteine (Hcy) (Hyperhomocysteinemia) (HHcy) has been reported in AMD. 1 year ago Rfjj Solutions LLC 1 year ago L & L Consulting LLC 2 years ago Mendo Westwinds LLC 2 years ago Chemlab Records LLC 2 years ago Timber Ridge Farm LLC 2 years ago All Tribal Networks LLC 2 years ago Marchi Estates LLC 2 years ago Mendo Reserve LLC 2 years ago Covelo Cannabis Company LLC 2 years ago Ramm Ranch LLC 2 years ago West Hills Property Maintenance and Consulting LLC 2 years ago Bcc Farms LLC 2 years ago Victoria's Farms LLC 2 years ago Anchor & Rose LLC 2 years ago Pearland Property LLC 2 years ago Slewfoot LLC 2 years ago The a Team, LLC 2 years ago Tegridy Too LLC 2 years ago Bulldog Farms L.L.C.Age-related macular degeneration (AMD) is a leading cause of vision loss. 1 year ago Happy Camper Cleaning Company LLC 1 year ago Disa Construction 1 year ago Round Valley Harvest LLC 1 year ago Wonderland Wellness LLC 1 year ago Old Boys LLC 1 year ago Purity LLC 1 year ago Cbjm Investments LLC 1 year ago The Queens Land LLC 1 year ago Fairbanks Farms, LLC 1 year ago Alex's All Around Hauling LLC 1 year ago Ar Agricultural LLC 1 year ago A&K Trucking and Equipment LLC 1 year ago Smith Towing Services, LLC 1 year ago M & T Kitchen, Inc.

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