May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Differential Expression of Caspases and Fas/FasL in Mouse Strain-Dependent Hyperoxia-Induced Vessel Regression
Author Affiliations & Notes
  • C. K. Chan
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
  • L. N. Pham
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
  • J. Zhou
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
  • Y. Ding
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
  • C. Spee
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
  • S. J. Ryan
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
  • D. R. Hinton
    Ophthalmology, The Arnold and Mabel Beckman Macular Research Center at the Doheny Eye Institute, Los Angeles, California
    Pathology, Keck School of Medicine of the University of Southern California, Los Angeles, California
  • Footnotes
    Commercial Relationships C.K. Chan, None; L.N. Pham, None; J. Zhou, None; Y. Ding, None; C. Spee, None; S.J. Ryan, None; D.R. Hinton, None.
  • Footnotes
    Support Arnold and Mabel Beckman Foundation; NIH Grants EY01545 and EY03040
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 2976. doi:
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      C. K. Chan, L. N. Pham, J. Zhou, Y. Ding, C. Spee, S. J. Ryan, D. R. Hinton; Differential Expression of Caspases and Fas/FasL in Mouse Strain-Dependent Hyperoxia-Induced Vessel Regression. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2976.

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

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Abstract

Purpose:: We investigated the heterogeneity of oxygen-induced retinal vessel regression.

Methods:: Using a mouse model of hyperoxia-induced vessel regression, strain-dependent vaso-obliteration was quantified among multiple strains of mice in flat-mount retinal specimens under darkfield microscopy and apoptosis ELISA. Co-localization of isolectin B4 and TUNEL staining allowed identification of endothelial specific apoptosis. Strain-dependent variations in ocular posterior pole gene expression of caspase-3, -8, and -9, and Fas and Fas ligand (FasL) were determined at 0-120 h hyperoxia by quantitative real time RT-PCR. Caspase activities were confirmed by substrate cleavage assays. Indirect ELISA of Fas/FasL protein expression correlated with mRNA expression.

Results:: The following rank order was determined for mouse strain-dependent hyperoxia-induced retinal vessel regression: CD-1 > C57BL/6J, DBA > A/J, 129S3/SvIM; DBA > AKR > 129S3/SvIM (Kruskall-Wallis p < 0.003). TUNEL staining revealed that hyperoxia-induced retinal vessel regression occurs via endothelial apoptosis. Following 24h and 120h hyperoxia, apoptosis ELISA was increased 1.6- and 1.3-fold in posterior poles of C57BL/6J mice compared with 129S3/SvIM strain (p<0.0001). Throughout hyperoxia, a 1.4 to1.9-fold increase in caspase-3 mRNA was observed in C57BL/6J mice compared to 129S3/SvIM (p < 0.0007). At 12 and 24 h hyperoxia, peak inter-strain differences of 1.5- and 1.3-fold increased caspase-9 expression respectively were also observed in C57BL/6J mice compared to 129S3/SvIM strain (p < 0.05). At 24 h hyperoxia, caspase activity between inbred strains correlated with mRNA expression. Fas mRNA expression increased from 72 h through 120 h hyperoxia reaching peak 2.5- and 3.5-fold increased levels in C57BL/6J and 129S3/SvIM mice respectively compared to that of 0 h hyperoxia (p < 0.01). In contrast, throughout hyperoxia, 129S3/SvIM mice expressed 1.7- to 2.4-fold increased FasL mRNA levels compared to C57BL/6J strain (p < 0.003). Using indirect protein ELISA at 72 h hyperoxia, while membrane-bound FasL protein was similarly expressed between C57BL/6J and 129S3/SvIM animals, soluble FasL protein was increased 1.9-fold in 129S3/SvIM mice compared to C57BL/6J animals at 72 h hyperoxia (p < 0.0008).

Conclusions:: Strain-dependent expression of apoptotic factors influences heterogeneity in vessel regression.

Keywords: vascular occlusion/vascular occlusive disease • apoptosis/cell death • retinal neovascularization 
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