title: Retinal ganglion cell (RGC) programmed necrosis contributes to ischemia-reperfusion-induced retinal damage.
abstract: Retinal ischemia-reperfusion (IR) injury remains a common cause of blindness and has a final pathway of retinal ganglion cell (RGC) death by apoptosis and necrosis. RGC apoptosis was intensively studied in IR injury, while RGC necrosis did not receive nearly enough consideration since it was viewed as an accidental and unregulated cellular event. However, there is evidence that necrosis, like apoptosis, can be implemented by a programmed mechanism. In this study, we tested the role of RGC programmed necrosis (necroptosis) in IR-induced retinal injury. We employed the mouse model of retinal IR injury for in vivo experiments. The oxygen and glucose deprivation (OGD) model was used as an IR model in vitro. Primary RGCs were isolated by an immunopanning technique. Necrostatin 1 (Nec1) was used to inhibit necroptosis in in vitro and in vivo experiments. The changes in gene expression were assessed by quantitative RT-PCR. The distribution of proteins in the retina and in RGC cultures was evaluated by immunohistochemistry and immunocytochemistry, respectively. Our data suggest that proteins (Ripk1 and Ripk3), which initiate necroptosis, were present in normal and ischemic RGCs. Treatment with Nec1 significantly reduced retinal damage after IR. Increased RGC survival and reduced RGC necrosis following OGD were observed in Nec1-treated cultures. We found significantly reduced expression of genes coding pro-inflammatory markers Il1b, Ccl5, Cxcl10, Nos2 and Cybb in Nec1-treated ischemic retinas. Thus, our findings suggest that RGC necroptosis contributes to retinal damage after IR through direct loss of cells and induction of associated inflammatory responses.
authors.affiliation: Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami Miller School of Medicine, Miami, FL, USA.
authors.forename: Galina
authors.initials: G
authors.lastname: Dvoriantchikova
authors.affiliation: Department of Biochemistry, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, MA, USA.
authors.forename: Alexei
authors.initials: A
authors.lastname: Degterev
authors.affiliation: Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami Miller School of Medicine, Miami, FL, USA; Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, FL, USA. Electronic address: divanov@med.miami.edu.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Ivanov
mesh_terms: D000818:Animals; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D004195:Disease Models, Animal; D019084:Fluorescent Antibody Technique, Indirect; D005786:Gene Expression Regulation / Q000502:physiology; D007093:Imidazoles / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D018836:Inflammation Mediators / Q000502:physiology; D008297:Male; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D009336:Necrosis; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000378:metabolism; D015427:Reperfusion Injury / Q000378:metabolism / Q000503:physiopathology* / Q000517:prevention & control; D012164:Retinal Diseases / Q000378:metabolism / Q000503:physiopathology* / Q000517:prevention & control; D055213:Retinal Pigment Epithelium / Q000378:metabolism / Q000473:pathology*
chemical_list: D007093:Imidazoles; D007211:Indoles; D018836:Inflammation Mediators; D012333:RNA, Messenger; C507699:necrostatin-1; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C506251:Ripk1 protein, mouse; C485615:Ripk3 protein, mouse
keywords: Ripk1; Ripk3; ischemia–reperfusion; necroptosis; necrostatin 1; retinal damage; retinal ganglion cells
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