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NF-B Activation in Light-Induced Retinal Degeneration in a Mouse Model

From the Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland.

PURPOSE. To investigate the modulation of nuclear factor (NF)-B in light-induced photoreceptor degeneration in a mouse model.

METHODS. Mice were exposed to intense green light. Light-induced activation of NF-B and its nuclear localization were studied by immunohistochemistry. The NF-B DNA–binding activity in the retinas after exposure to light was measured by electrophoretic mobility shift assay (EMSA). Nuclear transactivation of NF-B in the photoreceptor cells was determined by quantitative real-time (qRT)-PCR. The amount of NF-B p65 in the photoreceptor cells after exposure to light was assessed by Western blot analysis. To obtain more photoreceptor-specific information, microdissected photoreceptor cells were used in some studies.

RESULTS. By an immunohistochemical method, the perinuclear region of the photoreceptor cells was heavily labeled with an antibody to activated NF-B after a 1-hour exposure to light. Nuclear localization of NF-B in the photoreceptor nucleus was seen at 12 hours. In the experiments involving 3 hours of exposure to light followed by recovery in the dark, nuclear localization of NF-B was also noted after 12 hours’ recovery in the dark. During continuous exposure to light, the NF-B DNA–binding activity gradually increased and reached its maximum at 12 hours. There was an increase of NF-B p65 protein at 3 hours. The mRNA levels of IB were upregulated after 6 hours’ exposure to light.

CONCLUSIONS. Intense light activated NF-B in the photoreceptor cells in vivo, increased the NF-B DNA–binding activity, and increased the expression of mRNA of IB, a target gene of NF-B.

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