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

被引：19

作者：

Gilhooley, Michael J. ^{[1
,4
,5
]}

Hickey, Doron G. ^{[1
,3
]}

Lindner, Moritz ^{[1
,4
,6
]}

Palumaa, Teele ^{[2
]}

Hughes, Steven ^{[1
,2
]}

Peirson, Stuart N. ^{[2
]}

MacLaren, Robert E. ^{[1
,4
,5
]}

Hankins, Mark W. ^{[1
,2
]}

机构：

[1] Univ Oxford, Nuffield Dept Clin Neurosci, Nuffield Lab Ophthalmol, Oxford OX1 3RE, England

[2] Univ Oxford, Nuffield Dept Clin Neurosci, Sleep & Circadian Neurosci Inst, Oxford OX1 3RE, England

[3] Royal Victorian Eye & Ear Hosp, Melbourne, Vic 002, Australia

[4] Oxford Eye Hosp, Oxford OX3 9DU, England

[5] Moorfields Eye Hosp, London EC1V 2PD, England

[6] Philipps Univ, Inst Physiol & Pathophysiol, Dept Neurophysiol, Deutschhausstr 1-2, D-35037 Marburg, Germany

来源：

EXPERIMENTAL EYE RESEARCH | 2021年 / 207卷

基金：

英国生物技术与生命科学研究理事会; 英国惠康基金;

关键词：

Bipolar cell; Gene expression; Optogenetics; Inherited retinal degeneration; ECTOPIC EXPRESSION; MOUSE MODEL; RD1; MOUSE; VISION; TRANSDUCTION; RECEPTOR; BINDING; PROTEIN; PROTEOGLYCAN; MICROARRAYS;

D O I：

10.1016/j.exer.2021.108553

中图分类号：

R77 [眼科学];

学科分类号：

100212 ;

摘要：

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 Pde6brd1/rd1 selectively expressing an enhanced yellow fluorescent protein (EYFP) as a marker in ON-bipolar cells. Subsequent mRNA extraction allowed Illumina (R) 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.

引用

页数：11

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