title: Neuroprotective effects of C3 exoenzyme in excitotoxic retinopathy.
abstract: The purpose of this study is to evaluate the neuroprotective effects of C3 exoenzyme (C3) on N-methyl-d-aspartate (NMDA)-induced retinopathy in rats. C3 was expressed in Escherichia. coli and purified by affinity chromatography. Immunofluorescence was performed in NIH 3T3 cells treated with C3 to verify the cellular uptake of the protein. NMDA was injected intravitreally into rat eyes with or without C3. At various time points after injection, eyes were enucleated. Hematoxylin/eosin staining was performed on retina cross-sections for morphological analysis. Survival and apoptosis of cells in the ganglion cell layer (GCL) were assessed by cresyl violet staining and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) on retina flat-mounts. RhoA levels in retina cells were evaluated by Western blot to detect C3 uptake in vivo. The cellular uptake of C3 was verified by immunofluorescence. Damage including a decrease in inner plexiform layer (IPL) thickness and reduction of cell density in the GCL, corresponding to apoptosis of neurons, was induced by intravitreal injection of NMDA. Protection against this damage was observed following co-injection of C3 and NMDA. RhoA ADP-ribosylation induced by C3 was confirmed by Western blot. Our results suggest that C3 exerts neuroprotective effects against excitotoxic damage induced by NMDA.
authors.affiliation: Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Jinan University, Shenzhen, 518040, PR China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Wang
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Biotherapy, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, PR China.
authors.forename: Qiaona
authors.initials: Q
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Biotherapy, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, PR China.
authors.forename: Liheng
authors.initials: L
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Biotherapy, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yin
authors.affiliation: Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Jinan University, Shenzhen, 518040, PR China.
authors.forename: Ning
authors.initials: N
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Biotherapy, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, PR China.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Jinan University, Shenzhen, 518040, PR China.
authors.forename: Su-Ping
authors.initials: SP
authors.lastname: Cai
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Wisconsin, Madison, WI, USA.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Kaufman
authors.affiliation: Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Jinan University, Shenzhen, 518040, PR China. Electronic address: xliu1213@126.com.
authors.forename: Xuyang
authors.initials: X
authors.lastname: Liu
mesh_terms: D036002:ADP Ribose Transferases / Q000494:pharmacology*; D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001905:Botulinum Toxins / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D058449:Intravitreal Injections; D008297:Male; D051379:Mice; D016202:N-Methylaspartate; D041681:NIH 3T3 Cells; D018696:Neuroprotective Agents / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000187:drug effects*; D012164:Retinal Diseases / Q000139:chemically induced / Q000517:prevention & control*; D012165:Retinal Ganglion Cells / Q000187:drug effects; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D018696:Neuroprotective Agents; D016202:N-Methylaspartate; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins; D020742:rhoA GTP-Binding Protein
keywords: C3; excitotoxicity; glaucoma; neuroprotection