title: OFD1, as a Ciliary Protein, Exhibits Neuroprotective Function in Photoreceptor Degeneration Models.
abstract: Ofd1 is a newly identified causative gene for Retinitis pigmentosa (RP), a photoreceptor degenerative disease. This study aimed to examine Ofd1 localization in retina and further to investigate its function in photoreceptor degeneration models. Ofd1 localization in rat retina was examined using immunofluorescence. N-methyl-N-nitrosourea (MNU)-induced rats and Royal College of Surgeons (RCS) rats were used as photoreceptor degeneration models. The expression pattern of Ofd1, other ciliary associated genes and Wnt signaling pathway genes were examined in rat models. Furthermore, pEGFP-Ofd1-CDS and pSUPER-Ofd1-shRNA were constructed to overexpress and knockdown the expression level in 661W and R28 cells. MNU was also used to induce cell death. Cilia formation was observed using immunocytochemistry (ICC). Reactive oxygen species (ROS) were detected using the 2', 7'-Dichlorofluorescin diacetate (DCFH-DA) assay. Apoptosis genes expression was examined using qRT-PCR, Western blotting and fluorescence-activated cell sorting (FACS). Ofd1 localized to outer segments of rat retina photoreceptors. Ofd1 and other ciliary proteins expression levels increased from the 1st and 4th postnatal weeks and decreased until the 6th week in the RCS rats, while their expression consistently decreased from the 1st and 7th day in the MNU rats. Moreover, Wnt signaling pathway proteins expression was significantly up-regulated in both rat models. Knockdown of Ofd1 expression resulted in a smaller population, shorter length of cell cilia, and lower cell viability. Ofd1 overexpression partially attenuated MNU toxic effects by reducing ROS levels and mitigating apoptosis. To the best of our knowledge, this is the first study demonstrating Ofd1 localization and its function in rat retina and in retinal degeneration rat models. Ofd1 plays a role in controlling photoreceptor cilium length and number. Importantly, it demonstrates a neuroprotective function by protecting the photoreceptor from oxidative stress and apoptosis. These data have expanded our understanding of Ofd1 function beyond cilia, and we concluded that ofd1 neuroprotection could be a potential treatment strategy in retina degeneration models.
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Jieping
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Zongyi
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Jingying
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Furong
authors.initials: F
authors.lastname: Gao
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Caixia
authors.initials: C
authors.lastname: Jin
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Haibin
authors.initials: H
authors.lastname: Tian
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Jingfa
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Ophthalmology, Drexel University College of Medicine, Philadelphia, Pennsylvania.
authors.forename: Weiye
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.
authors.forename: Lixia
authors.initials: L
authors.lastname: Lu
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China.|Department of Regenerative Medicine and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, China.|Institute for Nutritional Sciences, Tongji University, Shanghai, China.
authors.forename: Guo-Tong
authors.initials: GT
authors.lastname: Xu
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis; D016923:Cell Death; D002460:Cell Line; D002469:Cell Separation; D002923:Cilia / Q000378:metabolism*; D000072661:Ciliopathies; D004596:Electroretinography; D005434:Flow Cytometry; D007150:Immunohistochemistry; D008770:Methylnitrosourea / Q000494:pharmacology; D051379:Mice; D018384:Oxidative Stress; D010786:Photoreceptor Cells / Q000473:pathology*; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011506:Proteins / Q000235:genetics*; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D012160:Retina / Q000378:metabolism / Q000473:pathology*; D012162:Retinal Degeneration / Q000378:metabolism / Q000473:pathology; D012174:Retinitis Pigmentosa / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology
chemical_list: D000975:Antioxidants; C544472:OFD1 protein, rat; D011506:Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D008770:Methylnitrosourea