May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Mechanisms of Macrophage–Induced Apoptosis in Mouse Retinal Pigment Epithelial Cells
Author Affiliations & Notes
  • D. Yang
    Ophthal & Vis Sci, Univ of Michigan–Kellogg Eye Ctr, Ann Arbor, MI
  • S.G. Elner
    Ophthal & Vis Sci, Univ of Michigan–Kellogg Eye Ctr, Ann Arbor, MI
  • L.–R. Lin
    Ophthal & Vis Sci, Univ of Michigan–Kellogg Eye Ctr, Ann Arbor, MI
  • V.N. Reddy
    Ophthal & Vis Sci, Univ of Michigan–Kellogg Eye Ctr, Ann Arbor, MI
  • V.M. Elner
    Ophthal & Vis Sci, Univ of Michigan–Kellogg Eye Ctr, Ann Arbor, MI
    Department of Pathology, University of Michigan, Ann Arbor, MI
  • Footnotes
    Commercial Relationships  D. Yang, None; S.G. Elner, None; L. Lin, None; V.N. Reddy, None; V.M. Elner, None.
  • Footnotes
    Support  NIH Grants EY09441 (VME), EY00484 (VNR) and EY07003 (core). V.M. Elner is a recipient of Lew R. Wasserman Merit Award from Research to Prevent Blindness.
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 1372. doi:
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    • Get Citation

      D. Yang, S.G. Elner, L.–R. Lin, V.N. Reddy, V.M. Elner; Mechanisms of Macrophage–Induced Apoptosis in Mouse Retinal Pigment Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1372.

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

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Abstract

Purpose: : Oxidative stress from reactive oxygen species (ROS) has been implicated in many diseases, including age–related macular degeneration (AMD), in which the retinal pigment epithelium (RPE) is considered a primary target. The purpose of this study was to determine whether macrophage–induced RPE apoptosis was involved in ROS and caspases.

Methods: : Mouse RPE cell cultures were established from wild–type (WT) and heterozygous manganese superoxide dismutase (Sod2)–knockout mouse (HET). The intracellular ROS, hydrogen peroxide (H2O2), and superoxide anion (O2), were measured by using 5–(and 6)–chloromethyl–2`,7`–dichlorodihydrofluorescence diacetate, acetyl ester (CM–H2DCFDA) and dihydroethidium (Het) assays, respectively. Apoptosis was evaluated by Hoechst staining and TUNEL assay. Changes in mitochondrial membrane potential were detected by JC–1 dye. Activated caspases were detected in situ by FITC–VAD–fmk staining.

Results: : IFN–γ–activated macrophages induced the production of intracellular ROS, decrease in mitochondrial membrane potential, activation of caspases and apoptosis in mRPE cells. The macrophage–induced ROS production, activation of caspases and apoptosis were significantly enhanced in the RPE cells derived from heterozygous Sod2–knockout mice.

Conclusions: : Our results suggest that activated macrophages may promote RPE cell apoptosis by inducing intracellular ROS production, and disruption of mitochondrial membrane potential and activation of caspases.

Keywords: retinal pigment epithelium • apoptosis/cell death • age-related macular degeneration 
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