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Alternative programs of cell death in developing retinal tissue.

Cinthya A Guimarães1, Marlene Benchimol, Gustavo P Amarante-Mendes

  • 1Instituto de Biofísica, Universidade Federal do Rio de Janeiro, CCS bloco G, Cidade Universitária, 21949-900 Rio de Janeiro, Brazil.

The Journal of Biological Chemistry
|August 15, 2003
PubMed
Summary
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Inhibition of protein synthesis triggers diverse cell death pathways in developing retinal cells, involving apoptosis and autophagy. Caspase-6 inhibition blocks TUNEL-negative cell death, revealing complex mechanisms.

Area of Science:

  • Cell Biology
  • Neuroscience
  • Developmental Biology

Background:

  • Protein synthesis inhibition is a known trigger for cell death in developing tissues.
  • Developing retinal cells undergo programmed cell death, involving apoptosis and autophagy.

Purpose of the Study:

  • To investigate the specific pathways of cell death induced by protein synthesis inhibition in developing retinal tissue.
  • To elucidate the roles of caspases and autophagy in these cell death processes.

Main Methods:

  • Developing retinal tissue was treated with protein synthesis inhibitors.
  • Ultrastructural analysis was performed.
  • Terminal dUTP nick-end labeling (TUNEL) and antibodies for activated caspases-3 and -9 were used.
  • Specific inhibitors of autophagy, caspases (3, 6, 9), and bongkrekic acid were employed.

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Main Results:

  • Protein synthesis inhibition induced both apoptosis and autophagy in retinal cells.
  • Only half of degenerating cells were TUNEL-positive or reacted with caspase-3/-9 antibodies.
  • Bongkrekic acid completely blocked cell death.
  • Inhibitors of autophagy, caspase-9, or -3 prevented only TUNEL-positive cell death.
  • Caspase-6 inhibition blocked TUNEL-negative cell death.
  • Simultaneous inhibition of caspases-9 and -6 nearly abolished cell death.

Conclusions:

  • Protein synthesis inhibition activates multiple, mitochondria-dependent cell death pathways in the developing retina.
  • Autophagy precedes caspase-9, caspase-3 activation, and DNA fragmentation.
  • Caspase-6 mediates a parallel, TUNEL-negative cell death pathway.
  • Distinct caspase inhibition can reveal additional cell death mechanisms.