Caspase activation in an experimental model of retinal detachment

Invest Ophthalmol Vis Sci. 2003 Mar;44(3):1262-7. doi: 10.1167/iovs.02-0492.

Abstract

Purpose: To test for apoptotic photoreceptor cell death and caspase activation as a function of time after induction of an experimental retinal detachment.

Methods: Retinal detachments were created in Brown Norway rats by injecting 10% hyaluronic acid into the subretinal space using a transvitreous approach. Light microscopy and terminal dUTP-biotin nick end-labeling (TUNEL) was performed at 1, 3, 5, and 7 days after detachment to assess for the morphologic features associated with apoptosis. Western blot analysis of retinal protein extracts was performed using antibodies against caspase-3, -7, and -9 and poly-ADP ribose-polymerase (PARP) at 1, 3, and 5 days after detachment.

Results: Light microscopic analysis of detached retinas showed the presence of pyknotic nuclei in the outer nuclear layer and disruption of the normal organization of the photoreceptor outer segments. TUNEL-staining was positive in the outer nuclear layer only in the detached portions of the retina. Western blot analysis confirmed the time-dependent activation of caspase-3, -7, and -9 and PARP in the detached retinas. No morphologic stigmata of apoptosis or caspase activation was detected in attached retinas.

Conclusions: The apoptotic photoreceptor cell death in experimental retinal detachments is associated with caspase activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Blotting, Western
  • Caspase 3
  • Caspase 7
  • Caspase 9
  • Caspases / metabolism*
  • Disease Models, Animal
  • Enzyme Activation
  • In Situ Nick-End Labeling
  • Male
  • Photoreceptor Cells, Vertebrate / enzymology*
  • Photoreceptor Cells, Vertebrate / pathology
  • Poly(ADP-ribose) Polymerases / metabolism
  • Rats
  • Rats, Inbred BN
  • Retinal Detachment / enzymology*
  • Retinal Detachment / pathology
  • Time Factors

Substances

  • Poly(ADP-ribose) Polymerases
  • Casp3 protein, rat
  • Casp9 protein, rat
  • Caspase 3
  • Caspase 7
  • Caspase 9
  • Caspases