June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Retinal pigment epithelium (RPE) response to selective retina therapy (SRT) in mouse eyes
Author Affiliations & Notes
  • Tae Kwann Park
    Ophthalmology, Soonchunhyang Univ Hospital, Bucheon-si, Korea (the Republic of)
  • Hoon Dong Kim
    Ophthalmology, Soonchunhyang Univ Hospital, Bucheon-si, Korea (the Republic of)
  • Ralf Brinkmann
    Medical Laser Center Lübeck GmbH, Lübeck, Germany
  • Young-Hoon Ohn
    Ophthalmology, Soonchunhyang Univ Hospital, Bucheon-si, Korea (the Republic of)
  • Footnotes
    Commercial Relationships Tae Kwann Park, None; Hoon Dong Kim, None; Ralf Brinkmann, None; Young-Hoon Ohn, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 5671. doi:
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    • Get Citation

      Tae Kwann Park, Hoon Dong Kim, Ralf Brinkmann, Young-Hoon Ohn; Retinal pigment epithelium (RPE) response to selective retina therapy (SRT) in mouse eyes. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5671.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

This study was designed to evaluate selective retina therapy (SRT)-RPE tissue reaction after SRT with 1.7 micro-second pulsed laser controlled by an automatic feedback reflectometry to avoid extended thermomechanical tissue damage.

 
Methods
 

Ten shots of SRT were performed in the right eyes of C57BL/6J mice using a Q-switched Nd:YLF laser system. SRT-treated mice received IP injection of 5-ethynyl-2'-deoxyuridine (EdU) in PBS. The mice were sacrificed at 3 hours to 14 days after treatment. Infrared (IR) and fluorescein angiographic (FA) images were taken with a confocal scanning laser ophthalmoscope (Heidelberg Retina Angiograph 2®; Heidelberg Engineering, Heidelberg, Germany). The whole mount and transverse sections were analyzed with In situ cell death detection kit, POD (Roche, Germany), Click-iT® Assay Kits (Life Technologies, Carlsbad, CA) for RPE cell proliferation, immunofluorescence (IF) staining with various antibodies. The changes of RPE cell numbers within 200 μm diameter centered at SRT site were counted at 3 hours to 14 days (n=5 at each time point). Untreated and conventional laser-treated mice were compared as negative and positive control.

 
Results
 

SRT sites were not detected with IR image but clearly detected with FA. At the SRT-treated area of the retinochoroidal tissue, only RPE cells were specifically detected with TUNEL (+) labeling. EdU (+) RPE cells were detected from 1 day after treatment, increased to 7 days and remained to 14 days. With whole mount sections, breakdown of cell-cell and expansion of RPE cells were detected by β-catenin IF staining. The number of RPE cells at SRT sites was gradually decreased to 12 hours (75.6% of baseline) and recovered to 14 days (93.4% of baseline). Up-regulated expression of Otx2 transcription factor was observed in the RPE cells located at SRT site.

 
Conclusions
 

SRT induced RPE cell death without photoreceptor cell damage. SRT-treated RPE areas were recovered with expansion and proliferation of the surrounding RPE cells.

 
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