HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (18) Citing Articles via Google Scholar Google Scholar Articles by Charles, I. Articles by Agarwal, N. Search for Related Content PubMed PubMed Citation Articles by Charles, I. Articles by Agarwal, N.

Serum Deprivation Induces Apoptotic Cell Death of Transformed Rat Retinal Ganglion Cells via Mitochondrial Signaling Pathways

¹From the Departments of Cell Biology and Genetics, University of North Texas Health Science Center, Fort Worth, Texas; and the ³Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, Kentucky.

PURPOSE. Apoptosis-related signaling pathways were investigated in a cultured rat retinal ganglion cell (RGC-5) line deprived of growth factors after serum withdrawal from the culture medium.

METHODS. RGC-5 cells were subjected to serum deprivation for 2 to 6 days and compared with RGC-5 cells cultured in growth medium containing 10% fetal calf serum. Cell viability was determined by a neutral red dye uptake assay. Apoptosis of RGC-5 cells was established by DNA laddering. The expression of various apoptosis-related genes was investigated by immunoblot analysis, and or reverse transcription polymerase chain reaction (RT-PCR) analysis. The redox state of the cell was determined by biochemical methods, including NF-B binding activity by electrophoretic mobility gel shift assays (EMSA) and mitochondrial damage by JC-1 (5,5', 6,6'-tetrachloro 1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide) staining, using live cell confocal microscopy and cytosolic release of cytochrome c.

RESULTS. Fifty percent cell loss was evident after 2 days of serum deprivation, as demonstrated by neutral red dye uptake assay. This cell loss was due to apoptotic cell death, as established by DNA laddering. The oxidative state of serum-deprived RGC-5 cells was perturbed as suggested by the increase in malonyldialdehyde (MDA) and a decrease in reduced glutathione (GSH) levels in cell lysates. The apoptosis of the RGC-5 cells was associated with the activation of caspase-3, -8, and -9, and increased levels of Bax with corresponding decreases in Bcl-2 levels and NF-B (NF-B) binding activity. Serum deprivation was also associated with a loss of mitochondrial function, as revealed by cytosolic release of cytochrome c and JC-1 staining of mitochondria of dying RGC-5 cells.

CONCLUSIONS. Taken together, these results indicate that serum withdrawal induces apoptotic cell death in RGC-5 cells via mitochondrial pathways. These studies lead to the speculation that growth factor deprivation arising from blockade of retrograde transport of neurotrophins may involve similar mechanism(s) of retinal ganglion cell death in glaucoma.

This article has been cited by other articles:

				S. K. Biswas, Y. Zhao, A. Nagalingam, T. W. Gardner, and L. Sandirasegarane PDGF- and Insulin/IGF-1-Specific Distinct Modes of Class IA PI 3-Kinase Activation in Normal Rat Retinas and RGC-5 Retinal Ganglion Cells Invest. Ophthalmol. Vis. Sci., August 1, 2008; 49(8): 3687 - 3698. [Abstract] [Full Text] [PDF]

				D. P. McKernan, M. B. Guerin, C. J. O'Brien, and T. G. Cotter A Key Role for Calpains in Retinal Ganglion Cell Death Invest. Ophthalmol. Vis. Sci., December 1, 2007; 48(12): 5420 - 5430. [Abstract] [Full Text] [PDF]

				W.-K. Ju, Q. Liu, K.-Y. Kim, J. G. Crowston, J. D. Lindsey, N. Agarwal, M. H. Ellisman, G. A. Perkins, and R. N. Weinreb Elevated Hydrostatic Pressure Triggers Mitochondrial Fission and Decreases Cellular ATP in Differentiated RGC-5 Cells Invest. Ophthalmol. Vis. Sci., May 1, 2007; 48(5): 2145 - 2151. [Abstract] [Full Text] [PDF]

				R. Harvey and S. K. Chintala Inhibition of Plasminogen Activators Attenuates the Death of Differentiated Retinal Ganglion Cells and Stabilizes Their Neurite Network In Vitro Invest. Ophthalmol. Vis. Sci., April 1, 2007; 48(4): 1884 - 1891. [Abstract] [Full Text] [PDF]

				X. Qi, A. S. Lewin, L. Sun, W. W. Hauswirth, and J. Guy Suppression of Mitochondrial Oxidative Stress Provides Long-term Neuroprotection in Experimental Optic Neuritis Invest. Ophthalmol. Vis. Sci., February 1, 2007; 48(2): 681 - 691. [Abstract] [Full Text] [PDF]

				M. Chwa, S. R. Atilano, V. Reddy, N. Jordan, D. W. Kim, and M. C. Kenney Increased Stress-Induced Generation of Reactive Oxygen Species and Apoptosis in Human Keratoconus Fibroblasts Invest. Ophthalmol. Vis. Sci., May 1, 2006; 47(5): 1902 - 1910. [Abstract] [Full Text] [PDF]

				F. B. Berry, F. Mirzayans, and M. A. Walter Regulation of FOXC1 Stability and Transcriptional Activity by an Epidermal Growth Factor-activated Mitogen-activated Protein Kinase Signaling Cascade J. Biol. Chem., April 14, 2006; 281(15): 10098 - 10104. [Abstract] [Full Text] [PDF]