June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Ethambutol-induced autophagy deregulation and cytotoxic effects in retinal ganglion cells
Author Affiliations & Notes
  • Shun-Ping Huang
    Ophthalmology, Tzu Chi General Hospital, Hualien, Taiwan
    Molecular Biology and Human Genetics, Tzu Chi University, Hualien, Taiwan
  • Shu-Fung Lin
    Ophthalmology, Tzu Chi General Hospital, Hualien, Taiwan
  • Rong-Kung Tsai
    Ophthalmology, Tzu Chi General Hospital, Hualien, Taiwan
  • Footnotes
    Commercial Relationships Shun-Ping Huang, None; Shu-Fung Lin, None; Rong-Kung Tsai, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1812. doi:
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    • Get Citation

      Shun-Ping Huang, Shu-Fung Lin, Rong-Kung Tsai; Ethambutol-induced autophagy deregulation and cytotoxic effects in retinal ganglion cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1812.

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

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Abstract

Purpose: To investigate the mechanism of cytotoxic effects of ethambutol in retinal ganglion cells

Methods: Cultured retinal ganglion cells (RGC-5) were exposed to 2 to 12 M ethambutol (EMB) and cell viability was quantified by MTS assay. Autophagy was studied in RGC-5 cells staining with LC3b,beclin-1 and p62. Lysosomes in cells were stained with Lysotracker and LAMP-1.

Results: Following exposure to EMB, RGC-5 cells developed cytosolic vacuoles within 1 h and underwent cell lysis within 24 h. The levels of LC3-II, beclin-1 and p62, as well as the number autophagic vacuoles (AVs), increased following EMB treatment, indicating that autophagy was activated. Protein kinase C-delta inhibitor, rottlerin, protected RGC-5 cells from EMB-induced cytoplasmic vacuoles formation and cell death. Rottlerin attenuated EMB-induced caspase 3 activation in RGC-5 cells.

Conclusions: EMB induced vacuole formation and cell death in RGC-5 cells. Initially, vacuoles developed from enlarged lysosomes, followed by activation the autophagy pathway and formation of LC-3 positive AVs. EMB-induced cytotoxic effects in RGC-5 cells are intimately correlated and regulated by the PKC delta signal pathway.

Keywords: 691 retina: proximal (bipolar, amacrine, and ganglion cells) • 714 signal transduction • 613 neuro-ophthalmology: optic nerve  
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