ON-bipolar cell gene expression during retinal degeneration: Implications for optogenetic visual restoration.

Autor: Gilhooley MJ; Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom; The Oxford Eye Hospital, Oxford, OX3 9DU, United Kingdom; Moorfields Eye Hospital, London, EC1V 2PD, United Kingdom., Hickey DG; Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom; Royal Victorian Eye and Ear Hospital, Melbourne, 002, Australia., Lindner M; Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom; The Oxford Eye Hospital, Oxford, OX3 9DU, United Kingdom; Institute of Physiology and Pathophysiology, Department of Neurophysiology, Philipps University, Deutschhausstrasse 1-2, Marburg, 35037, Germany., Palumaa T; Sleep and Circadian Neuroscience Institute, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom., Hughes S; Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom; Sleep and Circadian Neuroscience Institute, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom., Peirson SN; Sleep and Circadian Neuroscience Institute, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom., MacLaren RE; Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom; The Oxford Eye Hospital, Oxford, OX3 9DU, United Kingdom; Moorfields Eye Hospital, London, EC1V 2PD, United Kingdom., Hankins MW; Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom; Sleep and Circadian Neuroscience Institute, Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, OX1 3RE, United Kingdom. Electronic address: mark.hankins@eye.ox.ac.uk.
Jazyk: angličtina
Zdroj: Experimental eye research [Exp Eye Res] 2021 Jun; Vol. 207, pp. 108553. Date of Electronic Publication: 2021 Mar 31.
DOI: 10.1016/j.exer.2021.108553
Abstrakt: Purpose: Retinal bipolar cells survive even in the later stages of inherited retinal degenerations (IRDs) and so are attractive targets for optogenetic approaches to vision restoration. However, it is not known to what extent the remodelling that these cells undergo during degeneration affects their function. Specifically, it is unclear if they are free from metabolic stress, receptive to adeno-associated viral vectors, suitable for opsin-based optogenetic tools and able to propagate signals by releasing neurotransmitter.
Methods: Fluorescence activated cell sorting (FACS) was performed to isolate labelled bipolar cells from dissociated retinae of litter-mates with or without the IRD mutation Pde6b rd1/rd1 selectively expressing an enhanced yellow fluorescent protein (EYFP) as a marker in ON-bipolar cells. Subsequent mRNA extraction allowed Illumina® microarray comparison of gene expression in bipolar cells from degenerate to those of wild type retinae. Changes in four candidate genes were further investigated at the protein level using retinal immunohistochemistry over the course of degeneration.
Results: A total of sixty differentially expressed transcripts reached statistical significance: these did not include any genes directly associated with native primary bipolar cell signalling, nor changes consistent with metabolic stress. Four significantly altered genes (Srm2, Slf2, Anxa7 & Cntn1), implicated in synaptic remodelling, neurotransmitter release and viral vector entry had immunohistochemical staining colocalising with ON-bipolar cell markers and varying over the course of degeneration.
Conclusion: Our findings suggest relatively few gene expression changes in the context of degeneration: that despite remodelling, bipolar cells are likely to remain viable targets for optogenetic vision restoration. In addition, several genes where changes were seen could provide a basis for investigations to enhance the efficacy of optogenetic therapies.
(Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: