In Vivo Fluorescence Retinal Imaging Following AAV2-Mediated Gene Delivery in the Rat Retina

Lee, Joo Yong; Hwang, Yoonha; Kim, Ji Hyun; Kim, Yu Seun; Jung, Bok Kyoung; Kim, Pilhan; Lee, Heuiran

doi:10.1167/iovs.15-18862

Cited by 13 publications

(12 citation statements)

References 21 publications

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“…However, the CTB labelling was not specific to RGCs, with only 53% of CTB labelled cells being RGCs 30 . Other researchers have described AAV-mediated GFP gene transfer for cross-sectional 28 , 31 or longitudinal 32 fluorescence imaging of the retina, but there is no previously published work that has quantified RGC density from in vivo images or reported the specificity of GFP labelling to RGCs. Martin et al .…”

Section: Discussionmentioning

confidence: 99%

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

Smith

Chauhan

2018

Sci Rep

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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.

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Section: Discussionmentioning

confidence: 99%

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

Smith

Chauhan

2018

Sci Rep

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“…Using our custom-built confocal scanning laser retinal angiography system ( 50 ), in vivo fundus FA and ICGA of the CNV lesion were performed. Continuous-wave laser modules at 488 and 785 nm were used as excitation sources for fluorescein and ICG, respectively.…”

Section: Methodsmentioning

confidence: 99%

Tie2 activation promotes choriocapillary regeneration for alleviating neovascular age-related macular degeneration

et al. 2019

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“…During the entire imaging experiments, the body temperature was maintained at 37°C with a heating pad and temperature sensor (Kent Scientific). Intravital retina imaging used a custom-built multicolor video-rate laser-scanning confocal microscope modified for mouse retinas (53). Two scanning mirrors, a rotating polygonal mirror (Lincoln Laser) and a galvanometer-based scanning mirror (Cambridge Technology), were used to generate a 2-dimensional (2D) laser-scanning pattern, which was delivered to the sample through the relay lenses.…”

Section: Methodsmentioning

confidence: 99%

Sustained inflammation after pericyte depletion induces irreversible blood-retina barrier breakdown

Ogura¹,

Kurata²,

et al. 2017

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In the central nervous system, endothelial cells (ECs) and pericytes (PCs) of blood vessel walls cooperatively form a physical and chemical barrier to maintain neural homeostasis. However, in diabetic retinopathy (DR), the loss of PCs from vessel walls is assumed to cause breakdown of the blood-retina barrier (BRB) and subsequent vision-threatening vascular dysfunctions. Nonetheless, the lack of adequate DR animal models has precluded disease understanding and drug discovery. Here, by using an anti-PDGFRβ antibody, we show that transient inhibition of the PC recruitment to developing retinal vessels sustained EC-PC dissociations and BRB breakdown in adult mouse retinas, reproducing characteristic features of DR such as hyperpermeability, hypoperfusion, and neoangiogenesis. Notably, PC depletion directly induced inflammatory responses in ECs and perivascular infiltration of macrophages, whereby macrophage-derived VEGF and placental growth factor (PlGF) activated VEGFR1 in macrophages and VEGFR2 in ECs. Moreover, angiopoietin-2 (Angpt2) upregulation and Tie1 downregulation activated FOXO1 in PC-free ECs locally at the leaky aneurysms. This cycle of vessel damage was shut down by simultaneously blocking VEGF, PlGF, and Angpt2, thus restoring the BRB integrity. Together, our model provides new opportunities for identifying the sequential events triggered by PC deficiency, not only in DR, but also in various neurological disorders.

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In Vivo Fluorescence Retinal Imaging Following AAV2-Mediated Gene Delivery in the Rat Retina

Abstract: Citation: Lee JY, Hwang Y, Kim JH, et al. In vivo fluorescence retinal imaging following AAV2-mediated gene delivery in the rat retina. Invest

Cited by 13 publications

References 21 publications

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

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

Tie2 activation promotes choriocapillary regeneration for alleviating neovascular age-related macular degeneration

Sustained inflammation after pericyte depletion induces irreversible blood-retina barrier breakdown

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