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Activation of nuclear factor-kappaB during retinal degeneration in rd mice
Hui-Yang Zeng1, Mark O M Tso, Shenghan Lai
1Peking University Eye Center, Peking University Third Hospital, Beijing, China.
Purpose:
Transcription factors of the nuclear factor-kappa beta (NF-kappaB) family have been demonstrated to play an important role in the regulation of gene expression in the chronic neurodegenerative disorders. The aims of the current study were to investigate the alteration of NF-kappaB activity during retinal degeneration in rd mice and further explore its role in photoreceptor apoptosis.
Methods:
Activation of NF-kappaB and its nuclear translocation in the retina of rd mice at postnatal days (P) 8, 10, 12, 14, 16, 18, and 28 were studied by immunohistochemical analysis using NF-kappaB P65 antibody. The amount of NF-kappaB P65 protein and NF-kappaB DNA-binding activity in the whole retina were assessed by western blot analysis and gel shift analysis, respectively. Expression of NF-kappaB in microglial cells labeled with CD11b was determined by double labeling.
Results:
NF-kappaB P65 nuclear translocation and its DNA binding activity started to increase in the rd retina at P10 and reached a peak at P12. Expressions of P65 remained at high levels from P12 to P18. Double labeling of P65 with CD11 at P14 showed colocalization of P65 in the microglial cells in the outer nuclear layer.
Conclusions:
NF-kappaB was activated in the retinal degeneration of rd mice. NF-kappaB modulation may play a role in the retinal degeneration through microglial activation.
Insights
Nuclear factor-kappa beta (NF-kappaB) activation increases during retinal degeneration in rd mice, peaking at postnatal day 12. This suggests NF-kappaB modulation, potentially via microglial activation, plays a role in photoreceptor apoptosis.
Area of Science:
- Neuroscience
- Molecular Biology
- Ophthalmology
Background:
- Nuclear factor-kappa beta (NF-kappaB) is crucial for gene regulation in neurodegenerative diseases.
- Understanding NF-kappaB's role in retinal degeneration is vital for developing therapeutic strategies.
Purpose of the Study:
- To investigate alterations in NF-kappaB activity during retinal degeneration in rd mice.
- To explore the specific role of NF-kappaB in photoreceptor apoptosis.
Main Methods:
- Immunohistochemistry using NF-kappaB P65 antibody to assess nuclear translocation.
- Western blot and gel shift analysis to quantify NF-kappaB P65 protein and DNA-binding activity.
- Double labeling with CD11b to identify NF-kappaB expression in microglial cells.
Main Results:
- NF-kappaB P65 nuclear translocation and DNA-binding activity increased significantly from postnatal day 10, peaking at day 12.
- Elevated P65 expression persisted from day 12 to day 18.
- NF-kappaB P65 was found in microglial cells within the outer nuclear layer at day 14.
Conclusions:
- NF-kappaB is demonstrably activated during the process of retinal degeneration in rd mice.
- NF-kappaB modulation, possibly through microglial activation, is implicated in the progression of retinal degeneration.
