Time evolution of active caspase‐3 labelling after <i>in vivo</i> exposure to <scp>UVR</scp>‐300 nm

Talebizadeh, Nooshin; Yu, Zhu Liang; Kronschläger, Martin; Söderberg, Per G.

doi:10.1111/aos.12407

Cited by 8 publications

(7 citation statements)

References 22 publications

(32 reference statements)

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“…Ayala et al showed that double threshold dose of 8 kJ/m 2 of UVR-B increases mRNA and protein expression of both p53 and caspase 3 in the rat lens epithelium 7 d after exposure [18]. Moreover, a recent study in our group concluded that expression of active caspase 3 protein increases with a peak of 16 h after exposure to sub-threshold dose of 1 kJ/m 2 of UVR-B [19]. Galichanin et al investigated the time evolution of GADD45α, TP53 and CASP3 gene expression in the rat lens after in vivo exposure to double threshold dose of 8 kJ/m 2 of UVR-B [20].…”

Section: Introductionmentioning

confidence: 70%

Upregulation of GADD45a, TP53 and CASP3 mRNA Expression in the Rat Lens after In vivo Exposure to Sub-threshold Dose of UVR B

2014

J Ocul Biol

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The objective of the present study was to investigate the evolution of mRNA expression of the stress sensor GADD45α, the apoptosis initiator TP53 and the apoptosis executor CASP3 in the rat lens after in vivo exposure to sub-threshold dose of UVR-B.Methods: Forty six-week-old female albino Sprague-Dawley rats were unilaterally exposed to a sub-threshold dose, 1 kJ/m 2 (1.1 W/ m 2 for 15 min), of UVR (λ max = 300 nm). Anesthetized and dilated eyes were exposed to UVB radiation. The animals were sacrificed at 1, 5, 24 and 120 h post-exposure. mRNA expression of the GADD45α, TP53 and CASP3 genes in the lenses was measured by quantitative RT-PCR, and fold change in mRNA expression between exposed and unexposed lenses was calculated.Results: mRNA expression for GADD45α increased to a 1.2 fold change at 1 h after exposure and then returned to no change at 120 h. mRNA expression for TP53 increased with a regression coefficient of 0.04 h -1 to a maximum of 1.67 fold change. mRNA expression for CASP3 increased with a regression coefficient of 4.5 x10 -3 rel. units/h to a 1.46 fold change at 120 h after exposure. Conclusions:A sub-threshold in vivo exposure to UVR-B causes a transient upregulation of the stress sensor GADD45α at 1 h after exposure, a saturating upregulation of TP53 and a subsequent constant upregulation of CASP3 in the rat lens.

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Section: Introductionmentioning

confidence: 70%

Upregulation of GADD45a, TP53 and CASP3 mRNA Expression in the Rat Lens after In vivo Exposure to Sub-threshold Dose of UVR B

2014

J Ocul Biol

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“…UV light has been shown to cause apoptosis in lens cells and in whole lenses (21,22,(27)(28)(29) and because UV light exposure is associated with cataract formation (18 -20), we sought to determine the effect of UVA light on the subunits of the integrin ␣V␤5 receptor and on the process of lens cell phagocytosis of apoptotic debris. To test this, human SRA 01/04 lens epithelial cells were exposed to UVA light using a UVA LED array.…”

Section: Uva Treatment Of Lens Epithelial Cells Results In Loss Of Bomentioning

confidence: 99%

“…Most of the UV light transmitted to the lens is therefore UVA light; this is primarily absorbed by the anterior lens epithelium, and long term UVA exposure is believed to contribute to lens damage (24,83,84). UV light exposure of lens epithelial cells is associated with apoptosis of lens epithelial cells (21,22,(27)(28)(29) and human cataract formation (18 -20); however, the mechanisms used by lens epithelial cells to deal with the downstream effects of UV light and other insults, i.e. apoptosis and accumulation of apoptotic debris to ensure epithelial cell homeostasis and lens transparency, remain largely unknown.…”

Section: Discussionmentioning

confidence: 99%

“…A major consequence of UV light exposure to the lens is damage to the light-facing monolayer of anterior epithelial cells (21)(22)(23)(24)) that overlie and are required for the homeostasis of the organelle-free fiber cells that make up the lens core (11,25,26). UV light exposure has been shown to result in apoptosis of lens epithelial cells (21,22,(27)(28)(29), and damage to lens epithelial cells and their subcellular components results in cataract formation (22, 24, 30 -32). These data suggest the need for a lens mechanism to remove apoptotic debris generated upon UV light exposure to prevent epithelial cell death and thereby maintain lens transparency.…”

mentioning

confidence: 99%

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Integrin αVβ5-mediated Removal of Apoptotic Cell Debris by the Eye Lens and Its Inhibition by UV Light Exposure

Chauss

Brennan

Бакина

et al. 2015

Journal of Biological Chemistry

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Accumulation of apoptotic material is toxic and associated with cataract and other disease states. Identification of mechanisms that prevent accumulation of apoptotic debris is important for establishing the etiology of these diseases. The ocular lens is routinely assaulted by UV light that causes lens cell apoptosis and is associated with cataract formation. To date, no molecular mechanism for removal of toxic apoptotic debris has been identified in the lens. Vesicular debris within lens cells exposed to UV light has been observed raising speculation that lens cells themselves could act as phagocytes to remove toxic apoptotic debris. However, phagocytosis has not been confirmed as a function of the intact eye lens, and no mechanism for lens phagocytosis has been established. Here, we demonstrate that the eye lens is capable of phagocytizing extracellular lens cell debris. Using high throughput RNA sequencing and bioinformatics analysis, we establish that lens epithelial cells express members of the integrin ␣V␤5-mediated phagocytosis pathway and that internalized cell debris co-localizes with ␣V␤5 and with RAB7 and Rab-interacting lysosomal protein that are required for phagosome maturation and fusion with lysosomes. We demonstrate that the ␣V␤5 receptor is required for lens epithelial cell phagocytosis and that UV light treatment of lens epithelial cells results in damage to the ␣V␤5 receptor with concomitant loss of phagocytosis. These data suggest that loss of ␣V␤5-mediated phagocytosis by the eye lens could result in accumulation of toxic cell debris that could contribute to UV light-induced cataract formation.

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“…The rat was unilaterally exposed to UVR-300 nm as described elsewhere (16). After the exposure, both lenses were extracted and prepared for immunofluorescence detection of active caspase-3 as described above (Sample preparation).…”

Section: Application Of the Automatic Algorithmmentioning

confidence: 99%

Objective automated quantification of fluorescence signal in histological sections of rat lens

Talebizadeh

Hagström

et al. 2017

Cytometry Pt A

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Visual quantification and classification of fluorescent signals is the gold standard in microscopy. The purpose of this study was to develop an automated method to delineate cells and to quantify expression of fluorescent signal of biomarkers in each nucleus and cytoplasm of lens epithelial cells in a histological section. A region of interest representing the lens epithelium was manually demarcated in each input image. Thereafter, individual cell nuclei within the region of interest were automatically delineated based on watershed segmentation and thresholding with an algorithm developed in Matlab™. Fluorescence signal was quantified within nuclei, cytoplasms and juxtaposed backgrounds. The classification of cells as labelled or not labelled was based on comparison of the fluorescence signal within cells with local background. The classification rule was thereafter optimized as compared with visual classification of a limited dataset. The performance of the automated classification was evaluated by asking 11 independent blinded observers to classify all cells (n = 395) in one lens image. Time consumed by the automatic algorithm and visual classification of cells was recorded. On an average, 77% of the cells were correctly classified as compared with the majority vote of the visual observers. The average agreement among visual observers was 83%. However, variation among visual observers was high, and agreement between two visual observers was as low as 71% in the worst case. Automated classification was on average 10 times faster than visual scoring. The presented method enables objective and fast detection of lens epithelial cells and quantification of expression of fluorescent signal with an accuracy comparable with the variability among visual observers. © 2017 International Society for Advancement of Cytometry.

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Time evolution of active caspase‐3 labelling after in vivo exposure to UVR‐300 nm

Cited by 8 publications

References 22 publications

Upregulation of GADD45a, TP53 and CASP3 mRNA Expression in the Rat Lens after In vivo Exposure to Sub-threshold Dose of UVR B

Upregulation of GADD45a, TP53 and CASP3 mRNA Expression in the Rat Lens after In vivo Exposure to Sub-threshold Dose of UVR B

Integrin αVβ5-mediated Removal of Apoptotic Cell Debris by the Eye Lens and Its Inhibition by UV Light Exposure

Objective automated quantification of fluorescence signal in histological sections of rat lens

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