June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Differential effects of high glucose on connexin 43 hemichannels in compact versus sparsely grown retinal endothelial cells
Author Affiliations & Notes
  • Edward Bae
    Medicine and Ophthalmology, Boston University School of Medicine, Boston, MA
  • Dongjoon Kim
    Medicine and Ophthalmology, Boston University School of Medicine, Boston, MA
  • Jean X Jiang
    Biochemistry, University of Texas Health Science Center, San Antonio, TX
  • Sayon Roy
    Medicine and Ophthalmology, Boston University School of Medicine, Boston, MA
  • Footnotes
    Commercial Relationships Edward Bae, None; Dongjoon Kim, None; Jean Jiang, None; Sayon Roy, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4272. doi:
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    • Get Citation

      Edward Bae, Dongjoon Kim, Jean X Jiang, Sayon Roy; Differential effects of high glucose on connexin 43 hemichannels in compact versus sparsely grown retinal endothelial cells. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4272.

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

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Abstract

Purpose: To investigate whether high glucose condition alters connexin 43 (Cx43) hemichannel activity in retinal endothelial cells grown in sparse or dense condition and whether altered activity of Cx43 hemichannels influences apoptosis.

Methods: Rat retinal endothelial cells (RRECs) were grown in normal (5 mM glucose; N) or high glucose (30 mM; HG) medium for 7 days in either sparse or dense conditions. In parallel, cells grown in N or HG medium were exposed to a connexin channel blocker 18-β-glycyrrhetinic acid (GA) or or a non-active analogue of GA, glycyrrhizic acid (GZ) as control. To determine hemichannel activity, the dye uptake method was employed. Cells undergoing apoptosis was assessed using differential staining with acridine orange/ethidium bromide and TUNEL assay.

Results: Dye uptake results indicate RRECs grown in sparse condition exhibit increased hemichannel activity compared to those grown in densely populated cell condition. Interestingly, while HG tends to increase hemichannel activity in cells grown under sparsely populated condition, HG appeared to reduce hemichannel activity in densely populated cell condition. Importantly, when hemichannel activity was blocked apoptosis was observed, and at increasing concentrations of GA apoptosis was accelerated, whereas GZ had no effect. Thus, a biphasic effect of HG was observed as cells proliferated from a sparse to a dense, compact condition: an initial upregulation of hemichannel activity followed by sustained uncoupling effect.

Conclusions: Findings from this study indicate that Cx43 hemichannels play a distinctive role in response to HG in retinal endothelial cells in contrast to its role in gap junction intercellular communication activity. Importantly, increased hemichannel activity in the sparse cell populations under HG condition may signal a survival response countering HG-induced apoptosis in the pathogenesis of diabetic retinopathy.

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