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Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry
11:01

Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry

Published on: July 5, 2017

Cellular inhibitor of apoptosis (cIAP1) is down-regulated during retinal ganglion cell (RGC) maturation

Lilian Kisiswa1, Julie Albon, James E Morgan

  • 1School of Optometry and Vision Sciences, Cardiff University, Maindy Road, Cardiff, CF24 4LU Wales, UK.

Experimental Eye Research
|September 14, 2010
PubMed

Insights

As rats mature, retinal ganglion cells show reduced levels of anti-apoptotic proteins (IAPs), particularly cIAP1, and increased TRAF2. This suggests mature retinal cells may be more vulnerable to cell death.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Ophthalmology

Background:

  • Apoptosis, a key cell death mechanism, is implicated in aging and neurodegenerative diseases like glaucoma.
  • Understanding cell death regulation in the retina is crucial for developing treatments for vision loss.

Purpose of the Study:

  • To characterize the expression profiles of caspases (pro-apoptosis) and inhibitors of apoptosis (IAPs; anti-apoptosis) during the maturation of Brown Norway rat retina.
  • To examine changes in tumor necrosis factor receptor associated factor 2 (TRAF2) expression during retinal maturation.

Main Methods:

  • Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR were used to analyze gene expression.
  • Western blotting and immunofluorescence analysis were employed to assess protein levels and localization.

Main Results:

  • Caspase expression remained stable throughout retinal maturation.
  • A general reduction in IAP expression was observed, with a significant decrease in cellular IAP1 (cIAP1) in the retinal ganglion cell layer (RGCL).
  • Accumulation of TRAF2 was noted in the RGCL, correlating with cIAP1 down-regulation.

Conclusions:

  • Mature retinal ganglion cells may exhibit increased susceptibility to cell death due to reduced activation of survival pathways involving IAPs and TRAFs.
  • These findings provide insights into age-related changes in retinal cell vulnerability and potential mechanisms underlying glaucoma pathogenesis.

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