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Symantas Ragauskas, Sabine Grüner, Ludovic Collin, Giedrius Kalesnykas; Subretinal injection of poly(IC) causes acute photoreceptor cell death in the mouse retinal detachment model. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5377.
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© ARVO (1962-2015); The Authors (2016-present)
To determine the effect of poly(IC) subretinal injection on programmed retinal cell death.
Male 6-8 weeks old C57Bl/6J mice (n=30) were used. Subretinal injections were performed using poly(IC) (polyinosinicpolycytidylic acid/potassium salt) or phosphate buffer saline (PBS). The animals were followed for 2 days, 5 days and 7 days after subretinal injection. In vivo spectral domain-optical coherence tomography (SD-OCT) was employed to verify successful subretinal injection at the baseline. SD-OCT was also performed at the end of the follow-up period prior to animal sacrifice. The mice were transcardially perfused and the eyes were enucleated and cryosectioned. Ocular sections were systematically chosen and stained for hematoxylin and eosin (H&E) and for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The number of TUNEL-positive cells as well as the thickness of outer nuclear layer (ONL) were estimated using StereoInvestigator software (v. 10, MicroBrightField Inc., USA).
Both injections of PBS and poly(IC) caused retinal detachment. However, polyIC-injected group with a 2-day follow-up showed a significant increase in TUNEL-positive cells (5.2±1.48 cells/100 µm of ONL length, mean±SD) found in ONL as compared to other poly (IC) time-points (5-day group: 1.2±0.82 cells/100 µm of ONL length; 7-day group: 0.37±0.48 cells/100 µm of ONL length) or PBS-injected eyes (2-day group: 0.65±0.82 cells/100 µm of ONL length; 5-day group: 0.38±0.40 cells/100 µm of ONL length; 7-day group: 0.34±0.29 cells/100 µm of ONL length).
Subretinal injection of poly(IC) causes acute programmed cell death of photoreceptor cells. The use of SD-OCT is particularly important to verify and follow-up this preclinical model.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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