Effects of DJ-1 protein on retinal ganglion cells and visual function in mice after optic nerve crush injury_Chinese Journal of Ocular Fundus Diseases

Authors：

Ouyang Lingyi , He Tao ,  Xing Yiqiao

Eye Center of Renmin Hospital of Wuhan University, Wuhan 430060, China;

Corresponding author：

Xing Yiqiao, Email: yiqiao_xing57@whu.edu.cn

Keywords：

Optic nerve injuries; Retinal ganglion cells; Electroretinograph; Animal experimentation; DJ-1 protein

DOI：

10.3760/cma.j.cn511434-20201218-00621

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the effect of DJ-1 encoded by Park7 gene on retinal ganglion cells (RGC) and visual function after optic nerve crush injury (ONC) in mice.Methods Thirty-seven and 116 healthy male C57BL/6J mice were randomly divided into group normal, group ONC 2d, group ONC 5d, group ONC 7d and group control, group Park7, group Park7-ONC, group ONC and group green fluorescent protein (GFP)-ONC. Group ONC 2d, group ONC 5d and group ONC 7d were sacrificed on the 2nd, 5th and 7th day after the establishment of ONC model, and the follow-up experiments were carried out. The mice in group Park7 and group Park7-ONC were injected 1 μ recombinant adeno-associated virus (rAAV) with knocking down Park7 gene into vitreous cavity, and 1 μ l rAAV with only GFP was injected into vitreous cavity of mice in group GFP- ONC, and virus transfection was observed 4 weeks after injection. The injury of ONC was perfomed at 23 days after vitreous injection in group ONC, group Park7-ONC and group GFP-ONC, and the samples were taken for follow-up experiment 5 days after modeling. The average density of RGC was observed by immunofluorescence staining, the latencies and amplitudes of a-wave, b-wave and photopic negative response (phNR) and the amplitude of oscillatory potential (OPs)were detected by full-field flash electroretinogram,and the visual acuity of mice was measured by optomotor response (OMR). The relative expression levels of DJ-1, Bax and B lymphoblastoma / leukemia-2 (Bcl-2) protein in the retina of mice in each group were detected by Western blot. One-way ANOVA was used to compare the data between groups, and t-test was used for pairwise comparison between groups.Results Compared with the normal group, the relative expression of DJ-1 protein in the retina of the ONC 2 d group and ONC 5 d group increased significantly, and the difference was statistically significant (t=16.610, 5.628, P＜0.01,＜0.05). Four weeks after virus transfection, strong GFP expression was seen in the RGC layer and inner plexiform layer of the retina of mice in the Park7 group. Compared with the control group, the RGC density of the retina in the ONC group decreased significantly, and the difference was statistically significant (t=16.520, P＜0.000); compared with the ONC group, the RGC density of the retina in the Park7-ONC group decreased significantly, and the difference was statistically significant (t=6.074, P＜0.01). With the increase of stimulus light intensity, the dark adaptation a wave and b wave latency of the mice in the control group gradually shortened, and the amplitude gradually increased. The stimulus light intensity was 3 cd·s/m². There was no statistically significant difference in the dark adaptation a wave and b wave latency and amplitude of the control group, Park7 group, Park7-ONC group, ONC group, and GFP-ONC group (Incubation period: F=0.503, 2.592; P=0.734, 0.068. Amplitude: F=0.439, 1.451; P=0.779, 0.247). Compared with the control group, the Ops and PhNR amplitudes of the ONC group mice were significantly decreased (t=15.07, 12.80; P＜0.000,＜0.001). Compared with the ONC group, the Ops and PhNR amplitudes of the mice in the Park7-ONC group were significantly decreased (t=4.042, 5.062; P＜0.05,＜0.01); there was no statistically significant difference in the PhNR latency of the mice in each group (F=1.327, P=0.287). Compared with the control group, the visual acuity of the mice in the ONC group was significantly decreased, and the difference was statistically significant (t=23.020, P＜0.000); compared with the ONC group, the visual acuity of the mice in the Park7-ONC group was significantly decreased, and the difference was statistically significant (t=3.669, P＜0.05). Compared with the control group, Park7-ONC group and ONC group, the relative expression of DJ-1 protein in the mouse retina was significantly down-regulated, and the difference was statistically significant (t=47.140, 26.920; P＜0.000,＜0.000). There was no significant difference between ONC group and GFP-ONC group (t=0.739, P=0.983). Compared with the ONC group, the relative expression of Bax protein in the mouse retina of the Park7-ONC group was significantly increased, and the relative expression of Bcl-2 protein was significantly reduced. The differences were statistically significant (t=5.960, 9.710; P＜0.05,＜0.05); the relative expression ratio of Bcl-2/Bax in the Park7-ONC group was significantly lower than that in the ONC group, and the difference was statistically significant (t=13.620, P＜0.01).Conclusion The expression of DJ-1 encoded by Park7 gene is down-regulated after Park7 gene was knocked down, which aggravates the RGC damage and the decrease of retinal electrophysiological response and visual function in ONC injury mice.

Citation： Ouyang Lingyi, He Tao, Xing Yiqiao. Effects of DJ-1 protein on retinal ganglion cells and visual function in mice after optic nerve crush injury. Chinese Journal of Ocular Fundus Diseases, 2021, 37(5): 377-384. doi: 10.3760/cma.j.cn511434-20201218-00621 Copy

1.	Liu SA, Zhao ZN, Sun NN, et al. Transitions of the understanding and definition of primary glaucoma[J]. Chin Med J (Engl), 2018, 131(23): 2852-2859. DOI: 10.4103/0366-6999.246069.
2.	Weinreb RN, Aung T, Medeiros FA. The pathophysiology and treatment of glaucoma: a review[J]. JAMA, 2014, 311(18): 1901-1911. DOI: 10.1001/jama.2014.3192.
3.	He S, Stankowska DL, Ellis DZ, et al. Targets of neuroprotection in glaucoma[J]. J Ocul Pharmacol Ther, 2018, 34(1-2): 85-106. DOI: 10.1089/jop.2017.0041.
4.	Doozandeh A, Yazdani S. Neuroprotection in glaucoma[J]. J Ophthalmic Vis Res, 2016, 11(2): 209-220. DOI: 10.4103/2008-322X.183923.
5.	Yang RX, Lei J, Wang BD, et al. Pretreatment with sodium phenylbutyrate alleviates cerebral ischemia/reperfusion injury by upregulating DJ-1 protein[J/OL]. Front Neurol, 2017, 8: 256[2017-06-09]. https://pubmed.ncbi.nlm.nih.gov/28649223/. DOI: 10.3389/fneur.2017.00256.
6.	Bonifati V, Rizzu P, van Baren MJ, et al. Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism[J]. Science, 2003, 299(5604): 256-259. DOI: 10.1126/science.1077209.
7.	Ariga H, Takahashi-Niki K, Kato I, et al. Neuroprotective function of DJ-1 in Parkinson's disease[J/OL]. Oxid Med Cell Longev, 2013, 2013: 683920[2013-05-16]. https://pubmed.ncbi.nlm.nih.gov/23766857/. DOI: 10.1155/2013/683920.
8.	Gao H, Yang W, Qi Z, et al. DJ-1 protects dopaminergic neurons against rotenone-induced apoptosis by enhancing ERK-dependent mitophagy[J]. J Mol Biol, 2012, 423(2): 232-248. DOI: 10.1016/j.jmb.2012.06.034.
9.	Kitamura Y, Inden M, Kimoto Y, et al. Effects of a DJ-1-binding compound on spatial learning and memory impairment in a mouse model of Alzheimer's disease[J]. J Alzheimers Dis, 2017, 55(1): 67-72. DOI: 10.3233/JAD-160574.
10.	Satue M, Rodrigo MJ, Obis J, et al. Evaluation of progressive visual dysfunction and retinal degeneration in patients with Parkinson's disease[J]. Invest Ophthalmol Vis Sci, 2017, 58(2): 1151-1157. DOI: 10.1167/iovs.16-20460.
11.	Yun H, Lathrop KL, Yang E, et al. A laser-induced mouse model with long-term intraocular pressure elevation[J/OL]. PLoS One, 2014, 9(9): e107446[2014-09-12]. https://pubmed.ncbi.nlm.nih.gov/25216052/. DOI: 10.1371/journal.pone.0107446.
12.	Park SJ, Paik SS, Lee JY, et al. Blue-on-green flash induces maximal photopic negative response and oscillatory potential and serves as a diagnostic marker for glaucoma in rat retina[J]. Exp Neurobiol, 2018, 27(3): 210-216. DOI: 10.5607/en.2018.27.3.210.
13.	Kretschmer F, Sajgo S, Kretschmer V, et al. A system to measure the optokinetic and optomotor response in mice[J]. J Neurosci Methods, 2015, 256: 91-105. DOI: 10.1016/j.jneumeth.2015.08.007.
14.	刘一帆, 沈吟. 低浓度氯喹保护成年小鼠视网膜神经节细胞抵抗N-甲基-D-天冬氨酸的兴奋性毒性作用[J]. 中华眼底病杂志, 2020, 36(4): 295-301. DOI: 10.3760/cma.j.cn511434-20190903-00275.Liu YF, Sen Y. Low-dose chloroquine mediated neuroprotection against n-methyl-d-aspartate induced excitotoxicity in adult mice[J]. Chin J Ocul Fundus Dis, 2020, 36(4): 295-301. DOI: 10.3760/cma.j.cn511434-20190903-00275.
15.	Hijioka M, Inden M, Yanagisawa D, et al. DJ-1/PARK7: a new therapeutic target for neurodegenerative disorders[J]. Biol Pharm Bull, 2017, 40(5): 548-552. DOI: 10.1248/bpb.b16-01006.
16.	Oh SE, Mouradian MM. Regulation of signal transduction by DJ-1[J]. Adv Exp Med Biol, 2017, 1037: 97-131. DOI: 10.1007/978-981-10-6583-5_8.
17.	Taira T, Saito Y, Niki T, et al. DJ-1 has a role in antioxidative stress to prevent cell death[J]. EMBO Rep, 2004, 5(2): 213-218. DOI: 10.1038/sj.embor.7400074.
18.	Shadrach KG, Rayborn ME, Hollyfield JG, et al. DJ-1-dependent regulation of oxidative stress in the retinal pigment epithelium (RPE)[J/OL]. PLoS One, 2013, 8(7): e67983[2013-07-02]. https://pubmed.ncbi.nlm.nih.gov/23844142/. DOI: 10.1371/journal.pone.0067983.
19.	You Y, Gupta VK, Li JC, et al. Optic neuropathies: characteristic features and mechanisms of retinal ganglion cell loss[J]. Rev Neurosci, 2013, 24(3): 301-321. DOI: 10.1515/revneuro-2013-0003.
20.	Li Y, Cohen ED, Qian H. Rod and cone coupling modulates photopic ERG responses in the mouse retina[J/OL]. Front Cell Neurosci, 2020, 14: 566712[2020-09-25]. https://pubmed.ncbi.nlm.nih.gov/33100974/. DOI: 10.3389/fncel.2020.566712.
21.	Gauvin M, Dorfman AL, Trang N, et al. Assessing the contribution of the oscillatory potentials to the genesis of the photopic ERG with the discrete wavelet transform[J/OL]. Biomed Res Int, 2016, 2016: 2790194[2016-12-22]. https://pubmed.ncbi.nlm.nih.gov/28101507/. DOI: 10.1155/2016/2790194.
22.	Fairless R, Williams SK, Katiyar R, et al. ERG responses in mice with deletion of the synaptic ribbon component RIBEYE[J/OL]. Invest Ophthalmol Vis Sci, 2020, 61(5): 37[2020-05-11]. https://pubmed.ncbi.nlm.nih.gov/32437548/. DOI: 10.1167/iovs.61.5.37.
23.	Prencipe M, Perossini T, Brancoli G, et al. The photopic negative response (PhNR): measurement approaches and utility in glaucoma[J]. Int Ophthalmol, 2020, 40(12): 3565-3576. DOI: 10.1007/s10792-020-01515-0.
24.	Brayer S, Joannes A, Jaillet M, et al. The pro-apoptotic BAX protein influences cell growth and differentiation from the nucleus in healthy interphasic cells[J]. Cell Cycle, 2017, 16(21): 2108-2118. DOI: 10.1080/15384101.2017.1371882.
25.	Chen HC, Kanai M, Inoue-Yamauchi A, et al. An interconnected hierarchical model of cell death regulation by the BCL-2 family[J]. Nat Cell Biol, 2015, 17(10): 1270-1281. DOI: 10.1038/ncb3236.
26.	Maes ME, Schlamp CL, Nickells RW. BAX to basics: how the BCL2 gene family controls the death of retinal ganglion cells[J]. Prog Retin Eye Res, 2017, 57: 1-25. DOI: 10.1016/j.preteyeres.2017.01.002.
27.	Choi WS, Eom DS, Han BS, et al. Phosphorylation of p38 MAPK induced by oxidative stress is linked to activation of both caspase-8-and-9-mediated apoptotic pathways in dopaminergic neurons[J]. J Biol Chem, 2004, 279(19): 20451-20460. DOI: 10.1074/jbc.M311164200.
28.	Bode JG, Ehlting C, Häussinger D. The macrophage response towards LPS and its control through the p38(MAPK)-STAT3 axis[J]. Cell Signal, 2012, 24(6): 1185-1194. DOI: 10.1016/j.cellsig.2012.01.018.
29.	Pantcheva P, Elias M, Duncan K, et al. The role of DJ-1 in the oxidative stress cell death cascade after stroke[J]. Neural Regen Res, 2014, 9(15): 1430-1433. DOI: 10.4103/1673-5374.139458.
30.	Canet-Avilés RM, Wilson MA, Miller DW, et al. The Parkinson's disease protein DJ-1 is neuroprotective due to cysteine-sulfinic acid-driven mitochondrial localization[J]. Proc Natl Acad Sci USA, 2004, 101(24): 9103-9108. DOI: 10.1073/pnas.0402959101.
31.	Zhong N, Xu J. Synergistic activation of the human MnSOD promoter by DJ-1 and PGC-1alpha: regulation by SUMOylation and oxidation[J]. Hum Mol Genet, 2008, 17(21): 3357-3367. DOI: 10.1093/hmg/ddn230.
32.	Qu D, Rashidian J, Mount MP, et al. Role of Cdk5-mediated phosphorylation of Prx2 in MPTP toxicity and Parkinson's disease[J]. Neuron, 2007, 55(1): 37-52. DOI: 10.1016/j.neuron.2007.05.033.
33.	Wang W, Zhao H, Chen B. DJ-1 protects retinal pericytes against high glucose-induced oxidative stress through the Nrf2 signaling pathway[J/OL]. Sci Rep, 2020, 10(1): 2477[2020-02-12]. https://pubmed.ncbi.nlm.nih.gov/32051471/. DOI: 10.1038/s41598-020-59408-2.

1. Liu SA, Zhao ZN, Sun NN, et al. Transitions of the understanding and definition of primary glaucoma[J]. Chin Med J (Engl), 2018, 131(23): 2852-2859. DOI: 10.4103/0366-6999.246069.
2. Weinreb RN, Aung T, Medeiros FA. The pathophysiology and treatment of glaucoma: a review[J]. JAMA, 2014, 311(18): 1901-1911. DOI: 10.1001/jama.2014.3192.
3. He S, Stankowska DL, Ellis DZ, et al. Targets of neuroprotection in glaucoma[J]. J Ocul Pharmacol Ther, 2018, 34(1-2): 85-106. DOI: 10.1089/jop.2017.0041.
4. Doozandeh A, Yazdani S. Neuroprotection in glaucoma[J]. J Ophthalmic Vis Res, 2016, 11(2): 209-220. DOI: 10.4103/2008-322X.183923.
5. Yang RX, Lei J, Wang BD, et al. Pretreatment with sodium phenylbutyrate alleviates cerebral ischemia/reperfusion injury by upregulating DJ-1 protein[J/OL]. Front Neurol, 2017, 8: 256[2017-06-09]. https://pubmed.ncbi.nlm.nih.gov/28649223/. DOI: 10.3389/fneur.2017.00256.
6. Bonifati V, Rizzu P, van Baren MJ, et al. Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism[J]. Science, 2003, 299(5604): 256-259. DOI: 10.1126/science.1077209.
7. Ariga H, Takahashi-Niki K, Kato I, et al. Neuroprotective function of DJ-1 in Parkinson's disease[J/OL]. Oxid Med Cell Longev, 2013, 2013: 683920[2013-05-16]. https://pubmed.ncbi.nlm.nih.gov/23766857/. DOI: 10.1155/2013/683920.
8. Gao H, Yang W, Qi Z, et al. DJ-1 protects dopaminergic neurons against rotenone-induced apoptosis by enhancing ERK-dependent mitophagy[J]. J Mol Biol, 2012, 423(2): 232-248. DOI: 10.1016/j.jmb.2012.06.034.
9. Kitamura Y, Inden M, Kimoto Y, et al. Effects of a DJ-1-binding compound on spatial learning and memory impairment in a mouse model of Alzheimer's disease[J]. J Alzheimers Dis, 2017, 55(1): 67-72. DOI: 10.3233/JAD-160574.
10. Satue M, Rodrigo MJ, Obis J, et al. Evaluation of progressive visual dysfunction and retinal degeneration in patients with Parkinson's disease[J]. Invest Ophthalmol Vis Sci, 2017, 58(2): 1151-1157. DOI: 10.1167/iovs.16-20460.
11. Yun H, Lathrop KL, Yang E, et al. A laser-induced mouse model with long-term intraocular pressure elevation[J/OL]. PLoS One, 2014, 9(9): e107446[2014-09-12]. https://pubmed.ncbi.nlm.nih.gov/25216052/. DOI: 10.1371/journal.pone.0107446.
12. Park SJ, Paik SS, Lee JY, et al. Blue-on-green flash induces maximal photopic negative response and oscillatory potential and serves as a diagnostic marker for glaucoma in rat retina[J]. Exp Neurobiol, 2018, 27(3): 210-216. DOI: 10.5607/en.2018.27.3.210.
13. Kretschmer F, Sajgo S, Kretschmer V, et al. A system to measure the optokinetic and optomotor response in mice[J]. J Neurosci Methods, 2015, 256: 91-105. DOI: 10.1016/j.jneumeth.2015.08.007.
14. 刘一帆, 沈吟. 低浓度氯喹保护成年小鼠视网膜神经节细胞抵抗N-甲基-D-天冬氨酸的兴奋性毒性作用[J]. 中华眼底病杂志, 2020, 36(4): 295-301. DOI: 10.3760/cma.j.cn511434-20190903-00275.Liu YF, Sen Y. Low-dose chloroquine mediated neuroprotection against n-methyl-d-aspartate induced excitotoxicity in adult mice[J]. Chin J Ocul Fundus Dis, 2020, 36(4): 295-301. DOI: 10.3760/cma.j.cn511434-20190903-00275.
15. Hijioka M, Inden M, Yanagisawa D, et al. DJ-1/PARK7: a new therapeutic target for neurodegenerative disorders[J]. Biol Pharm Bull, 2017, 40(5): 548-552. DOI: 10.1248/bpb.b16-01006.
16. Oh SE, Mouradian MM. Regulation of signal transduction by DJ-1[J]. Adv Exp Med Biol, 2017, 1037: 97-131. DOI: 10.1007/978-981-10-6583-5_8.
17. Taira T, Saito Y, Niki T, et al. DJ-1 has a role in antioxidative stress to prevent cell death[J]. EMBO Rep, 2004, 5(2): 213-218. DOI: 10.1038/sj.embor.7400074.
18. Shadrach KG, Rayborn ME, Hollyfield JG, et al. DJ-1-dependent regulation of oxidative stress in the retinal pigment epithelium (RPE)[J/OL]. PLoS One, 2013, 8(7): e67983[2013-07-02]. https://pubmed.ncbi.nlm.nih.gov/23844142/. DOI: 10.1371/journal.pone.0067983.
19. You Y, Gupta VK, Li JC, et al. Optic neuropathies: characteristic features and mechanisms of retinal ganglion cell loss[J]. Rev Neurosci, 2013, 24(3): 301-321. DOI: 10.1515/revneuro-2013-0003.
20. Li Y, Cohen ED, Qian H. Rod and cone coupling modulates photopic ERG responses in the mouse retina[J/OL]. Front Cell Neurosci, 2020, 14: 566712[2020-09-25]. https://pubmed.ncbi.nlm.nih.gov/33100974/. DOI: 10.3389/fncel.2020.566712.
21. Gauvin M, Dorfman AL, Trang N, et al. Assessing the contribution of the oscillatory potentials to the genesis of the photopic ERG with the discrete wavelet transform[J/OL]. Biomed Res Int, 2016, 2016: 2790194[2016-12-22]. https://pubmed.ncbi.nlm.nih.gov/28101507/. DOI: 10.1155/2016/2790194.
22. Fairless R, Williams SK, Katiyar R, et al. ERG responses in mice with deletion of the synaptic ribbon component RIBEYE[J/OL]. Invest Ophthalmol Vis Sci, 2020, 61(5): 37[2020-05-11]. https://pubmed.ncbi.nlm.nih.gov/32437548/. DOI: 10.1167/iovs.61.5.37.
23. Prencipe M, Perossini T, Brancoli G, et al. The photopic negative response (PhNR): measurement approaches and utility in glaucoma[J]. Int Ophthalmol, 2020, 40(12): 3565-3576. DOI: 10.1007/s10792-020-01515-0.
24. Brayer S, Joannes A, Jaillet M, et al. The pro-apoptotic BAX protein influences cell growth and differentiation from the nucleus in healthy interphasic cells[J]. Cell Cycle, 2017, 16(21): 2108-2118. DOI: 10.1080/15384101.2017.1371882.
25. Chen HC, Kanai M, Inoue-Yamauchi A, et al. An interconnected hierarchical model of cell death regulation by the BCL-2 family[J]. Nat Cell Biol, 2015, 17(10): 1270-1281. DOI: 10.1038/ncb3236.
26. Maes ME, Schlamp CL, Nickells RW. BAX to basics: how the BCL2 gene family controls the death of retinal ganglion cells[J]. Prog Retin Eye Res, 2017, 57: 1-25. DOI: 10.1016/j.preteyeres.2017.01.002.
27. Choi WS, Eom DS, Han BS, et al. Phosphorylation of p38 MAPK induced by oxidative stress is linked to activation of both caspase-8-and-9-mediated apoptotic pathways in dopaminergic neurons[J]. J Biol Chem, 2004, 279(19): 20451-20460. DOI: 10.1074/jbc.M311164200.
28. Bode JG, Ehlting C, Häussinger D. The macrophage response towards LPS and its control through the p38(MAPK)-STAT3 axis[J]. Cell Signal, 2012, 24(6): 1185-1194. DOI: 10.1016/j.cellsig.2012.01.018.
29. Pantcheva P, Elias M, Duncan K, et al. The role of DJ-1 in the oxidative stress cell death cascade after stroke[J]. Neural Regen Res, 2014, 9(15): 1430-1433. DOI: 10.4103/1673-5374.139458.
30. Canet-Avilés RM, Wilson MA, Miller DW, et al. The Parkinson's disease protein DJ-1 is neuroprotective due to cysteine-sulfinic acid-driven mitochondrial localization[J]. Proc Natl Acad Sci USA, 2004, 101(24): 9103-9108. DOI: 10.1073/pnas.0402959101.
31. Zhong N, Xu J. Synergistic activation of the human MnSOD promoter by DJ-1 and PGC-1alpha: regulation by SUMOylation and oxidation[J]. Hum Mol Genet, 2008, 17(21): 3357-3367. DOI: 10.1093/hmg/ddn230.
32. Qu D, Rashidian J, Mount MP, et al. Role of Cdk5-mediated phosphorylation of Prx2 in MPTP toxicity and Parkinson's disease[J]. Neuron, 2007, 55(1): 37-52. DOI: 10.1016/j.neuron.2007.05.033.
33. Wang W, Zhao H, Chen B. DJ-1 protects retinal pericytes against high glucose-induced oxidative stress through the Nrf2 signaling pathway[J/OL]. Sci Rep, 2020, 10(1): 2477[2020-02-12]. https://pubmed.ncbi.nlm.nih.gov/32051471/. DOI: 10.1038/s41598-020-59408-2.

Chinese Journal of Ocular Fundus Diseases

Effects of DJ-1 protein on retinal ganglion cells and visual function in mice after optic nerve crush injury

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content