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Open AccessArticle

Degenerated Cones in Cultured Human Retinas Can Successfully Be Optogenetically Reactivated

1
Netherlands Institute for Neuroscience, 1105 BA Amsterdam-Zuidoost, The Netherlands
2
Department of Ophthalmology, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands
3
Department of Cell and Chemical Biology, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands
4
Department of Biomedical Engineering & Physics, Amsterdam University Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(2), 522; https://doi.org/10.3390/ijms21020522
Received: 5 December 2019 / Revised: 11 January 2020 / Accepted: 13 January 2020 / Published: 14 January 2020
(This article belongs to the Special Issue Retinal Degeneration: From Pathophysiology to Therapeutic Approaches)
Biblical references aside, restoring vision to the blind has proven to be a major technical challenge. In recent years, considerable advances have been made towards this end, especially when retinal degeneration underlies the vision loss such as occurs with retinitis pigmentosa. Under these conditions, optogenetic therapies are a particularly promising line of inquiry where remaining retinal cells are made into “artificial photoreceptors”. However, this strategy is not without its challenges and a model system using human retinal explants would aid its continued development and refinement. Here, we cultured post-mortem human retinas and show that explants remain viable for around 7 days. Within this period, the cones lose their outer segments and thus their light sensitivity but remain electrophysiologically intact, displaying all the major ionic conductances one would expect for a vertebrate cone. We optogenetically restored light responses to these quiescent cones using a lentivirus vector constructed to express enhanced halorhodopsin under the control of the human arrestin promotor. In these ‘reactivated’ retinas, we show a light-induced horizontal cell to cone feedback signal in cones, indicating that transduced cones were able to transmit their light response across the synapse to horizontal cells, which generated a large enough response to send a signal back to the cones. Furthermore, we show ganglion cell light responses, suggesting the cultured explant’s condition is still good enough to support transmission of the transduced cone signal over the intermediate retinal layers to the final retinal output level. Together, these results show that cultured human retinas are an appropriate model system to test optogenetic vision restoration approaches and that cones which have lost their outer segment, a condition occurring during the early stages of retinitis pigmentosa, are appropriate targets for optogenetic vision restoration therapies. View Full-Text
Keywords: cultured human retina; cone photoreceptors; optogenetics; retinitis pigmentosa; vision restoration cultured human retina; cone photoreceptors; optogenetics; retinitis pigmentosa; vision restoration
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Kamar, S.; Howlett, M.H.C.; Klooster, J.; de Graaff, W.; Csikós, T.; Rabelink, M.J.W.E.; Hoeben, R.C.; Kamermans, M. Degenerated Cones in Cultured Human Retinas Can Successfully Be Optogenetically Reactivated. Int. J. Mol. Sci. 2020, 21, 522.

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