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Zhang Y, Yin R, Liu X. Changes in Cyclic Guanosine Monophosphate Channel of 661w Cells In vitro with Excessive Light Time. JOVR [Internet]. 2023Nov.30 [cited 2024May14];18(4):417–423. Available from: https://knepublishing.com/index.php/JOVR/article/view/14554
https://doi.org/10.18502/jovr.v18i4.14554
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authors
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Yahan Zhang
Yahan Zhang
Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University, National Clinical Research Center for Eye Diseases, Shanghai Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China
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Rui Yin
Rui Yin
Department of Biomedical Informatics, Harvard Medical School, Boston, MA, USA
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Xin Liu
Xin Liu
Department of Ophthalmology, The Second Hospital of Jilin University, Changchun, China
Published Date
- Nov 30, 2023
Changes in Cyclic Guanosine Monophosphate Channel of 661w Cells In vitro with Excessive Light Time
Abstract
Purpose: To determine the response time and protective mechanism of the cyclic guanosine monophosphate (cGMP) channel in 661w cells.
Methods: 661w cells were exposed to 4500Lux visible light for three and four days at the following exposure time periods per day: 20, 30, 60, 90, 120, and 180. Cells were incubated for the rest of the time without any other treatment. Cell activity and cell death rates were measured with Hoechst/PI (diphenylmethane/propidium iodide) staining. Western Blot was used to detect the levels of guanylate cyclase-activating proteins 1 (GCAP1), cGMP, and phosphodiesterase (PDE)6 in the cGMP-gated channel.
Results: 661w cells showed low mortality within three days. The mortality rate increased from the fourth day, especially during the longer times (120 and 180 min) of light exposure. After three-day illumination, the level of cGMP increased after 20 and 90 min and the level of GCAP1 increased after 60 and 90 min. After four days of illumination, the level of GCAP1 upregulated after a time of 20 and 60 min, while the cGMP level decreased from 30 min. The expression of PDE6 upregulated at each light period.
Conclusion: The survival rate of 661w cells was relevant to the time of light exposure. The changes in GCAP1, cGMP, and PDE6 levels over time were possibly related to cell metabolism and restoration after light-induced damage.
Keywords:
Retinal Light-induced Injury, cGMP-gated Channel, Photoreceptor Cells
References
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22. Campbell LJ, Jensen AM. Phosphodiesterase inhibitors sildenafil and vardenafil reduce zebrafish rod photoreceptor outer segment shedding. Invest Ophthalmol Vis Sci 2017;58:5604–5615.
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26. Wyse Jackson AC, Cotter TG. The synthetic progesterone Norgestrel is neuroprotective in stressed photoreceptorlike cells and retinal explants, mediating its effects via basic fibroblast growth factor, protein kinase A and glycogen synthase kinase 3β signalling. Eur J Neurosci 2016;43:899–911.
27. Li GY, Li T, Fan B. The D₁ dopamine receptor agonist, SKF83959, attenuates hydrogen peroxide-induced injury in RGC-5 cells involving the extracellular signal-regulated kinase/p38 pathways. Mol Vis 2012;18:2882–2895.
28. Yan J, Chen Y, Zhu Y. Programmed non-apoptotic cell death in hereditary retinal degeneration: Crosstalk between cGMP-dependent pathways and PARthanatos? Int J Mol Sci 2021;22:10567.
29. Yan J, Günter A, Das S. Inherited retinal degeneration: PARP-dependent activation of Calpain Requires CNG channel activity. Biomolecules 2022;12:455.
30. Li Y, Li R, Dai H. Novel variants in PDE6A and PDE6B genes and its phenotypes in patients with retinitis pigmentosa in Chinese families. BMC Ophthalmol 2022;22:27.
2. Paskowitz DM, LaVail MM, Duncan JL. Light and inherited retinal degeneration. Br J Ophthalmol 2006;90:1060– 1066.
3. Zhao Y, Shen Y. Light-induced retinal ganglion cell damage and the relevant mechanisms. Cell Mol Neurobiol 2020;40:1243–1252.
4. Rose K, Walston ST, Chen J. Separation of photoreceptor cell compartments in mouse retina for protein analysis. Mol Neurodegeneration 2017;12:28.
5. Remé CE. The dark side of light: Rhodopsin and the silent death of vision. Invest Ophthalmol Vis Sci 2005;46:2672– 2682.
6. Kayatz P, Heimann K, Schraermeyer U. Ultrastructural localization of light-induced lipid peroxides in the rat retina. Invest Ophthalmol Vis Sci 1999;40:2314–2321.
7. Stryer, L. Cyclic GMP cascade of vision. Annu Rev Neurosci 1986;9:87–119.
8. Field GD, Uzzell V, Chichilnisky EJ, Rieke F. Temporal resolution of single-photon responses in primate rod photoreceptors and limits imposed by cellular noise. J Neurophysiol 2019;121:255—268.
9. Cote RH. Photoreceptor phosphodiesterase (PDE6): Activation and inactivation mechanisms during visual transduction in rods and cones. Pflugers Arch 2021;473:1377–1391.
10. Ames JB. Structural insights into retinal guanylate cyclase activator proteins (GCAPs). Int J Mol Sci 2021;22:8731.
11. Dizhoor AM, Olshevskaya EV, Peshenko IV. Mg2+/Ca2+ cation binding cycle of guanylyl cyclase activating proteins (GCAPs): Role in regulation of photoreceptor guanylyl cyclase. Mol Cell Biochem 2010334:117–124.
12. Tan E, Ding XQ, Saadi A, Agarwal N, Naash MI, Al-Ubaidi MR. Expression of cone-photoreceptor-specific antigens in a cell line derived from retinal tumors in transgenic mice. Invest Ophthalmol Vis Sci 2004;45:764–768.
13. Ebrey T, Koutalos Y. Vertebrate photoreceptors. Prog Retin Eye Res 2001;20:49-94.
14. Imanishi Y, Li N, Sokal I, Sowa ME, Lichtarge O, Wensel TG, Saperstein DA, Baehr W, Palczewski K. Characterization of retinal guanylate cyclase-activating protein 3 (GCAP3) from zebrafish to man. Eur J Neurosci 2002;15:63–78.
15. Johnson EC, Bacigalupo J. Spontaneous activity of the light-dependent channel irreversibly induced in excised patches from Limulus ventral photoreceptors. J Membr Biol 1992;130:33–47.
16. Li GY, Fan B, Jiao YY. Rapamycin attenuates visible lightinduced injury in retinal photoreceptor cells via inhibiting endoplasmic reticulum stress. Brain Res 2014;1563:1–12.
17. Wood JP, Osborne NN. Induction of apoptosis in cultured human retinal pigmented epithelial cells: The effect of protein kinase C activation and inhibition. Neurochem Int 1997;31:261–273.
18. Makino CL, Wen XH, Olshevskaya EV, Peshenko IV, Savchenko AB, Dizhoor AM. Enzymatic relay mechanism stimulates cyclic GMP synthesis in rod photoresponse: Biochemical and physiological study in guanylyl cyclase activating protein 1 knockout mice. PLoS One 2012;7:e47637.
19. Chen WJ, Wu C, Xu Z, Kuse Y, Hara H, Duh EJ. Nrf2 protects photoreceptor cells from photo-oxidative stress induced by blue light. Exp Eye Res 2017;154:151–158.
20. Dizhoor AM, Hurley JB. Regulation of photoreceptor membrane guanylyl cyclases by guanylyl cyclase activator proteins. Methods 1999;19:521–531.
21. Gao Y, Eskici G, Ramachandran S, Poitevin F, Seven AB, Panova O, et al. Structure of the visual signaling complex between transducin and phosphodiesterase 6. Mol Cell 2021;81:2496. Erratum for: Mol Cell 2020;80:237–245.e4.
22. Campbell LJ, Jensen AM. Phosphodiesterase inhibitors sildenafil and vardenafil reduce zebrafish rod photoreceptor outer segment shedding. Invest Ophthalmol Vis Sci 2017;58:5604–5615.
23. Cote RH. Photoreceptor phosphodiesterase (PDE6): Activation and inactivation mechanisms during visual transduction in rods and cones. Pflugers Arch 2021;473:1377–1391.
24. Iribarne M, Masai I. Do cGMP levels drive the speed of photoreceptor degeneration? Adv Exp Med Biol 2018;1074:327–333.
25. Organisciak DT, Vaughan DK. Retinal light damage: Mechanisms and protection. Prog Retin Eye Res 2010;29:113–134.
26. Wyse Jackson AC, Cotter TG. The synthetic progesterone Norgestrel is neuroprotective in stressed photoreceptorlike cells and retinal explants, mediating its effects via basic fibroblast growth factor, protein kinase A and glycogen synthase kinase 3β signalling. Eur J Neurosci 2016;43:899–911.
27. Li GY, Li T, Fan B. The D₁ dopamine receptor agonist, SKF83959, attenuates hydrogen peroxide-induced injury in RGC-5 cells involving the extracellular signal-regulated kinase/p38 pathways. Mol Vis 2012;18:2882–2895.
28. Yan J, Chen Y, Zhu Y. Programmed non-apoptotic cell death in hereditary retinal degeneration: Crosstalk between cGMP-dependent pathways and PARthanatos? Int J Mol Sci 2021;22:10567.
29. Yan J, Günter A, Das S. Inherited retinal degeneration: PARP-dependent activation of Calpain Requires CNG channel activity. Biomolecules 2022;12:455.
30. Li Y, Li R, Dai H. Novel variants in PDE6A and PDE6B genes and its phenotypes in patients with retinitis pigmentosa in Chinese families. BMC Ophthalmol 2022;22:27.