Assessment of Rat and Mouse RGC Apoptosis Imaging<i>in Vivo</i>with Different Scanning Laser Ophthalmoscopes

Maass, Annelie; Leithner, Peter Lundh von; Luong, Vy; Guo, Li; Salt, Thomas E.; Fitzke, F.W.; Cordeiro, M. Francesca

doi:10.1080/02713680701585872

Cited by 60 publications

(56 citation statements)

References 51 publications

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“…The ability to image fluorescent labeling of RGCs through the use of specialty confocal scanning laser microscopy, although not widely available, has been previously reported in both mice and rats (15). The activatable nature of the TcapQ probe, with quenching of fluorescence in its unactivated state, is advantageous in that it results in negligible background fluorescence, and thus enhanced signal-to-noise ratios.…”

Section: Discussionmentioning

confidence: 99%

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Single-cell imaging of retinal ganglion cell apoptosis with a cell-penetrating, activatable peptide probe in an in vivo glaucoma model

Barnett

Zhang

Maxwell

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

126

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Molecular imaging probes have potential for in vivo identification of apoptosis and other intracellular processes. TcapQ, a cell-penetrating, near-infrared fluorescent peptide probe designed to be optically silent through intramolecular fluorescence quenching and activated by effector caspases, has been previously described and validated in vitro. Herein, using NMDA-induced apoptosis of retinal ganglion cells (RGCs), representing an in vivo rat model of glaucoma, we assessed the ability of TcapQ to image single-cell apoptosis through effector caspase activity. Following intravitreal injection, intracellular TcapQ activation occurred specifically in RGCs, identified individual apoptotic cells, showed a clear dose-response relationship with NMDA, and colocalized with TUNEL labeling in the retina. There was a significant diminution of probe activation following pretreatment with a specific inhibitor of caspase-3. Stereospecificity was also exhibited by the lack of intracellular fluorescence upon administration of the noncleavable isomer, d TcapQ. TcapQ has potential utility in detecting and monitoring single-cell apoptosis in glaucoma in vivo.

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Section: Discussionmentioning

confidence: 99%

“…This strategy has been used to identify apoptotic RGCs in vivo (10,(14)(15)(16). One potential drawback to this strategy is that it may not adequately distinguish apoptotic from necrotic forms of cell death in vivo without a secondary marker of membrane integrity.…”

mentioning

confidence: 99%

Single-cell imaging of retinal ganglion cell apoptosis with a cell-penetrating, activatable peptide probe in an in vivo glaucoma model

Barnett

Zhang

Maxwell

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

126

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“…24,25 Although DARC will soon be tested for the first time in a glaucoma clinical trial, (ISRCTN59484478), all studies so far have used experimental models. 1,[11][12][13][14]26,27 For imaging, wide-field retina images are acquired and assessed for fluorescence using a method we have previously described. 28 The total number of apoptosing RGCs for each time point in vivo is then calculated (Figure 1), 13 to create a 'DARC count'.…”

Section: The Darc Technologymentioning

confidence: 99%

“…This has meant, for example, imaging with a wide-angle lens (Figure 1), the selection of different wavelengths to enhance signal-to-noise, and improving the correlation of the in vivo and histology counts. 13,51 As hypothesized in Table 1, it is anticipated that DARC will provide a snapshot of the number of apoptosing RGCs at any one timeFgiving rise to a DARC count. Large population-based studies will be needed to establish the DARC count in relation to glaucoma and the normal ageing process, to validate the estimates above.…”

Section: How Safe and Invasive Is The Technology?mentioning

confidence: 99%

“…9 The DARC Technology (Detection of Apoptosing Retinal Cells) is an innovative technique that uses the unique optical properties of the eye to allow direct visualization of nerve cells dying through apoptosis, identified by fluorescent-labeled annexin V. DARC has been used in various experimental models for the in vivo analysis of RGC apoptosis. 1,[10][11][12][13] Recent studies suggest its potential not only in investigating the pathogenesis of glaucoma, but also in its early diagnosis, as single cell resolution may enable the detection of abnormalities in the previously regarded 'sub-clinical' stages of the disease. Furthermore, the successful use of DARC in glaucoma models to assess the neuroprotective effects of potential drugs, 11,12,14,15 suggests that the technique may also be useful in patients as a tool to gauge therapeutic efficacy.…”

Section: Introductionmentioning

confidence: 99%

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Imaging apoptosis in the eye

Cordeiro

Migdal

Bloom

et al. 2011

Eye

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Apoptosis is a form of programmed cell death that is implicated in both pathological and physiological processes throughout the body. Its imaging in vivo with intravenous radiolabelled-annexin V has been heralded as an important advance, with around 30 clinical trials demonstrating its application in the early detection and monitoring of disease, and the assessment of efficacy of potential and existing therapies. A recent development has been the use of fluorescently labeled annexin V to visualize single retinal cells undergoing the process of apoptosis in vivo with ophthalmoscopy. This has been given the acronym DARC (Detection of Apoptosing Retinal Cells). DARC so far has only been used experimentally, but clinical trials are starting shortly in glaucoma patients. Results suggest that DARC may provide a direct assessment of retinal ganglion cell health. By enabling early assessment and quantitative analysis of cellular degeneration in glaucoma, it is hoped that DARC can identify patients before the onset of irreversible vision loss. Furthermore, in addition to aiding the tracking of disease, it may provide a rapid and objective assessment of potential and effective therapies, providing a new and meaningful clinical endpoint in glaucomatous disease that is so badly needed.

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Real-Time Imaging of Retinal Cell Apoptosis by Confocal Scanning Laser Ophthalmoscopy

Normando

Dehabadi

Guo

et al. 2014

Methods in Molecular Biology

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Retinal cell apoptosis occurs in many eye conditions, including glaucoma, diabetic retinopathy and Alzheimer's disease. Real-time detection of retinal cell apoptosis has potential clinical value in early disease detection, as well as evaluating disease progression and treatment efficacy. Here, we describe our novel imaging technology DARC (Detection of Apoptosing Retinal Cells), which can be used to visualize single retinal neurons undergoing apoptosis in real time, by using fluorescently labeled Annexin A5 and confocal scanning laser ophthalmoscopy (cSLO ). Clinical trials of DARC in glaucoma patients are due to start shortly, but in this chapter, we describe this technique in experimental animal models.

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Assessment of Rat and Mouse RGC Apoptosis Imagingin Vivowith Different Scanning Laser Ophthalmoscopes

Cited by 60 publications

References 51 publications

Single-cell imaging of retinal ganglion cell apoptosis with a cell-penetrating, activatable peptide probe in an in vivo glaucoma model

Single-cell imaging of retinal ganglion cell apoptosis with a cell-penetrating, activatable peptide probe in an in vivo glaucoma model

Imaging apoptosis in the eye

Real-Time Imaging of Retinal Cell Apoptosis by Confocal Scanning Laser Ophthalmoscopy

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