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J Biol Chem, Vol. 274, Issue 6, 3734-3743, February 5, 1999
B via Involvement of Caspase-1, Leading to Apoptosis of
Photoreceptor Cells
,,
From the Department of Anatomy and Cell Biology, North Texas Eye
Research Institute at University of North Texas Health Science Center,
Fort Worth, Texas 76107, the § Department of Pediatrics,
Louisiana State University Medical Center, Shreveport, Louisiana 71130, the The mechanisms of photoreceptor cell death via
apoptosis, in retinal dystrophies, are largely not understood. In the
present report we show that visible light exposure of mouse cultured
661W photoreceptor cells at 4.5 milliwatt/cm2 caused
a significant increase in oxidative damage of 661W cells, leading to
apoptosis of these cells. These cells show constitutive expression of
nuclear factor- Department of Molecular Oncology,
Cytokine Research Laboratory, University of Texas M. D. Anderson
Cancer Center, Houston, Texas 77030, and the ¶ Department of
Ophthalmology and Visual Sciences, University of Illinois at Chicago,
Chicago, Illinois 60612
B (NF-B), and light exposure of photoreceptor
cells results in lowering of NF-B levels in both the nuclear and
cytosolic fractions in a time-dependent manner. Immunoblot
analysis of IB
and p50, and p65 (RelA) subunits of NF-B,
suggested that photo-oxidative stress results in their depletion.
Immunocytochemical studies using antibody to RelA subunit of NF-B
further revealed the presence of this subunit constitutively both in
the nucleus and cytoplasm of the 661W cells. Upon exposure to
photo-oxidative stress, a depletion of the cytoplasmic and nuclear RelA
subunit was observed. The depletion of NF-B appears to be mediated
through involvement of caspase-1. Furthermore, transfection of these
cells with a dominant negative mutant IB greatly enhanced the
kinetics of down modulation of NF-B, resulting in a faster
photo-oxidative stress-induced apoptosis. Taken together, these studies
show that the presence of NF-B RelA subunit in the nucleus is
essential for protection of photoreceptor cells against apoptosis
mediated by an oxidative pathway.
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