In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells

Abstract A defining characteristic of optic neuropathies, such as glaucoma, is progressive loss of retinal ganglion cells (RGCs). Current clinical tests only provide weak surrogates of RGC loss, but the possibility of optically visualizing RGCs and quantifying their rate of loss could represent a ra...

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Autores principales:	Corey A. Smith, Balwantray C. Chauhan
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0bad900fd8e442df8a3cb462d6cc5e81
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spelling	oai:doaj.org-article:0bad900fd8e442df8a3cb462d6cc5e812021-12-02T15:08:58ZIn vivo imaging of adeno-associated viral vector labelled retinal ganglion cells10.1038/s41598-018-19969-92045-2322https://doaj.org/article/0bad900fd8e442df8a3cb462d6cc5e812018-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-19969-9https://doaj.org/toc/2045-2322Abstract A defining characteristic of optic neuropathies, such as glaucoma, is progressive loss of retinal ganglion cells (RGCs). Current clinical tests only provide weak surrogates of RGC loss, but the possibility of optically visualizing RGCs and quantifying their rate of loss could represent a radical advance in the management of optic neuropathies. In this study we injected two different adeno-associated viral (AAV) vector serotypes in the vitreous to enable green fluorescent protein (GFP) labelling of RGCs in wild-type mice for in vivo and non-invasive imaging. GFP-labelled cells were detected by confocal scanning laser ophthalmoscopy 1-week post-injection and plateaued in density at 4 weeks. Immunohistochemical analysis 5-weeks post-injection revealed labelling specificity to RGCs to be significantly higher with the AAV2-DCX-GFP vector compared to the AAV2-CAG-GFP vector. There were no adverse functional or structural effects of the labelling method as determined with electroretinography and optical coherence tomography, respectively. The RGC-specific positive and negative scotopic threshold responses had similar amplitudes between control and experimental eyes, while inner retinal thickness was also unchanged after injection. As a positive control experiment, optic nerve transection resulted in a progressive loss of labelled RGCs. AAV vectors provide strong and long-lasting GFP labelling of RGCs without detectable adverse effects.Corey A. SmithBalwantray C. ChauhanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Corey A. Smith Balwantray C. Chauhan In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
description	Abstract A defining characteristic of optic neuropathies, such as glaucoma, is progressive loss of retinal ganglion cells (RGCs). Current clinical tests only provide weak surrogates of RGC loss, but the possibility of optically visualizing RGCs and quantifying their rate of loss could represent a radical advance in the management of optic neuropathies. In this study we injected two different adeno-associated viral (AAV) vector serotypes in the vitreous to enable green fluorescent protein (GFP) labelling of RGCs in wild-type mice for in vivo and non-invasive imaging. GFP-labelled cells were detected by confocal scanning laser ophthalmoscopy 1-week post-injection and plateaued in density at 4 weeks. Immunohistochemical analysis 5-weeks post-injection revealed labelling specificity to RGCs to be significantly higher with the AAV2-DCX-GFP vector compared to the AAV2-CAG-GFP vector. There were no adverse functional or structural effects of the labelling method as determined with electroretinography and optical coherence tomography, respectively. The RGC-specific positive and negative scotopic threshold responses had similar amplitudes between control and experimental eyes, while inner retinal thickness was also unchanged after injection. As a positive control experiment, optic nerve transection resulted in a progressive loss of labelled RGCs. AAV vectors provide strong and long-lasting GFP labelling of RGCs without detectable adverse effects.
format	article
author	Corey A. Smith Balwantray C. Chauhan
author_facet	Corey A. Smith Balwantray C. Chauhan
author_sort	Corey A. Smith
title	In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
title_short	In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
title_full	In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
title_fullStr	In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
title_full_unstemmed	In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
title_sort	in vivo imaging of adeno-associated viral vector labelled retinal ganglion cells
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/0bad900fd8e442df8a3cb462d6cc5e81
work_keys_str_mv	AT coreyasmith invivoimagingofadenoassociatedviralvectorlabelledretinalganglioncells AT balwantraycchauhan invivoimagingofadenoassociatedviralvectorlabelledretinalganglioncells
_version_	1718387918793342976

In vivo imaging of adeno-associated viral vector labelled retinal ganglion cells

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