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Specific spatial distribution of caspase-3 in normal lenses
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Ophthalmology.ORCID iD: 0000-0001-7325-7358
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Ophthalmology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Ophthalmology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Developmental Neuroscience.
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2014 (English)In: Acta Ophthalmologica, ISSN 1755-375X, E-ISSN 1755-3768, Vol. 93, no 3, 289-292 p.Article in journal (Refereed) Published
Abstract [en]

Purpose

To determine the distribution of active caspase-3 in rat eye lens epithelium.

Methods

In total, 120 sagittal sections from forty rats were assessed for active caspase-3 labelling using immunohistochemistry. Lens epithelial cells were counted, and the fraction of active caspase-3 labelled cells and their relative positions were identified in each section.

Results

Active caspase-3 is present in normal lens epithelium. The active caspase-3 expression was higher in the anterior pole of the lens. Probability of radial spatial distribution of labelling was fitted with a logistic model. The increase rate and the inflection point were estimated as CI (0.95) to 23 ± 3 cells and 114 ± 3 cells, respectively.

Conclusion

The gradually decreasing active caspase-3 labelling from the anterior pole to the periphery suggests that active caspase-3 may be involved in normal protein turnover caused by, for example, incident light.

Place, publisher, year, edition, pages
2014. Vol. 93, no 3, 289-292 p.
National Category
Ophthalmology
Research subject
Ophtalmology
Identifiers
URN: urn:nbn:se:uu:diva-226542DOI: 10.1111/aos.12501ISI: 000353053000031OAI: oai:DiVA.org:uu-226542DiVA: diva2:726383
Available from: 2014-06-18 Created: 2014-06-18 Last updated: 2017-12-05Bibliographically approved
In thesis
1. Caspase-3 in lens epithelium
Open this publication in new window or tab >>Caspase-3 in lens epithelium
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Purpose: To model the time evolution of active caspase-3 protein expression in a healthy lens, and in a lens exposed to UVR-300 nm (UVR-B). To develop an automated method to classify the fluorescent signal of biomarkers in the lens epithelial cells.

Methods: Six-week old Sprague-Dawley rats were used. Firstly, expression of active caspase-3 was studied in the lens epithelium of healthy rats. Secondly, rats were unilaterally exposed in vivo to 1 kJ/m2 UVR-B for 15 minutes. At 0.5, 8, 16, and 24 hours after the UVR-B exposure, the exposed and the contralateral non-exposed lenses were removed. Immunohistochemistry was done on three mid-sagittal sections from each lens. The florescent labelling for active caspase-3 in each lens section was counted three times. The time evolution of active caspase-3 expression in response to UVR-B exposure was modelled as a function of cell position in the lens epithelium. An automated objective method was developed to quantify the lens epithelial cells and to classify the fluorescent signal of active caspase-3. Active caspase-3 was selected as a model signal.

Results: Active caspase-3 was abundant in the anterior pole of the normal lenses. Spatial distribution of active caspase-3 labelling in the lens epithelium was fitted to a logistic model. The probability of active caspase-3 expression was higher in the UVR-B exposed lenses (95% CI = 0.12 ± 0.01). There was no difference in the expression of active caspase-3 between the 0.5 and the 24 hours groups or between the 8 and the 16 hours groups. A difference was noted, when comparing the 0.5 and 24 hours groups with the 8 and 16 hours groups (Test statistic 7.01, F1;36;0.95= 4.11). Exposure to UVR-B has an impact on the average probability of labelling for active caspase-3 as a function of cell position. The probability of labelling as a function of cell number also varied as a function of time after UVR-B exposure. The automated method counted the lens epithelial cells and estimated the proportion of active caspase-3 labelling in the lens epithelium.

Conclusions: Active caspase-3 is present in the healthy lens epithelial cells. Active caspase-3 exhibits higher expression at the anterior pole of the lens and the expression decreases towards the periphery. After UVR-B exposure, the expression of active caspase-3 in the lens epithelium increases with a peak of expression occurring around 16 hours after exposure. The average probability of labelling in the lens epithelium is dependent on both the UVR-B exposure and the time period elapsed after the exposure. The automated method enables objective and fast quantification of lens epithelial cells and the expression of fluorescent signal in the lens cells.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2016. 40 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1168
Keyword
ultraviolet radiation, caspase-3, lens, cataract, apoptosis, Immunohistochemistry, spatial distribution, time evolution, modelling, automatic analysis, cell counting, image analysis.
National Category
Neurosciences
Research subject
Ophtalmology
Identifiers
urn:nbn:se:uu:diva-267543 (URN)978-91-554-9436-0 (ISBN)
Public defence
2016-02-05, Enghoffsalen, entrance 50, 1st floor, Akademiska Sjukhuset, Uppsala, 13:00 (English)
Opponent
Supervisors
Available from: 2016-01-21 Created: 2015-11-24 Last updated: 2016-02-12Bibliographically approved

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Publisher's full texthttp://onlinelibrary.wiley.com/doi/10.1111/aos.12501/pdf

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Talebizadeh, NooshinYu, ZhaohuaMartin, KronschlägerFinn, HallböökPer, Söderberg

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