June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Real-time in vivo detection of retinal reactive oxygen species
Author Affiliations & Notes
  • Ning Fan
    Shenzhen Eye Hospital,Jinan University, Shenzhen, China
    North Texas Eye Research Institute, Department of Pharmaceutical Sciences, UNT Health Science Center, Fort Worth, TX
  • Yang Liu
    North Texas Eye Research Institute, Department of Pharmaceutical Sciences, UNT Health Science Center, Fort Worth, TX
  • Byung-Jin Kim
    North Texas Eye Research Institute, Department of Pharmaceutical Sciences, UNT Health Science Center, Fort Worth, TX
  • Liping Tang
    Department of Bioengineering, University of Texas at Arlington, Arlington, TX
  • Abbot F Clark
    Cell Biology and Immunology,North Texas Eye Research Institute, UNT Health Science Center, Fort Worth, TX
  • Iok-Hou Pang
    North Texas Eye Research Institute, Department of Pharmaceutical Sciences, UNT Health Science Center, Fort Worth, TX
  • Footnotes
    Commercial Relationships Ning Fan, None; Yang Liu, None; Byung-Jin Kim, None; Liping Tang, None; Abbot Clark, Genzyme-Sonofi (C), ISIS Pharmaceuticals (C), Reata Pharmaceutical (F), Sanofi-Fovea (C); Iok-Hou Pang, Alcon (C), Novartis (C), Regeneron (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1995. doi:
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    • Get Citation

      Ning Fan, Yang Liu, Byung-Jin Kim, Liping Tang, Abbot F Clark, Iok-Hou Pang; Real-time in vivo detection of retinal reactive oxygen species. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1995.

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

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Abstract

Purpose: We characterized and optimized the use of a chemiluminescent probe L-012 as a non-invasive in vivomethod to assessoptic nerve crush(ONC)-induced accumulation of reactive oxygen species (ROS) in the mouseretina.

Methods: We performed ONC unilaterally in adult C57BL/6J mice to induce retinal damage. L-012-dependent luminescencein the eye was acquired in a small animalin vivo imaging system. Optimal detection time and dose of L-012 were empirically determined.Histology of the retina was performed by H&E staining of cross-sections.Protein carbonyl content in retina was measured byenzyme-linked immunosorbent assay.

Results: Ocular L-012 chemiluminescence was observed at Day 1 and Day 3 in the ONC-injured eye, but not in the contralateral uninjured control orsham surgery eye. The signal was dependent on the dose of L-012; 75mg/kg of L-012 providedhighest signalcompared with that of 25mg/kg or 7.5mg/kg.The peak chemiluminescence occurredbetween15min and20 min after intraperitoneal L-012injection (Peak photon radiance of ONC eye = 120 ± 61 x 103 p/s/cm2/sr;control eye = 8 ± 1 x 103 p/s/cm2/sr; mean ± SD, n = 5; p < 0.05).The signal correlated with oxidative changes in the retina: retinal protein carbonyl content was 1.36±0.16nmol/mg proteinat Day 1 after ONC, which was significantly (p < 0.05) higher than that ofthe control retina(0.51±0.05nmol/mg). Thechemiluminescence signal of Day 3 was significantly(p < 0.05) lower than that of Day 1.Intraperitoneal treatment with tempol (100mg/kg), a ROS scavenger, or apocynin (50mg/kg), a NADPH oxidase inhibitor significantly(p < 0.05) reduced ocular L-012 chemiluminescence. Histological evaluation of H&E stained retina cross-sections showed no evidence of retinal toxicity by L-012.

Conclusions: Currently available detection methods of ROS require post-mortem, ex vivo tissue or cell samples. Our data show thatL-012is a useful non-invasive in vivo tool to assess the accumulation of ROS in the retina. Thisreal-time in vivo imaging technology can significantly aid our understanding of the involvement of oxidative damage in various retinal injuries.

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