Comparison of the signal transduction pathways for the induction of gene expression of nitric oxide synthase-2 in response to two different stimuli
Section snippets
Human optic nerve head astrocyte cultures
Twelve human eyes from donors (aged 22–49 years) with no history of eye disease were obtained within 24 h after death from eye banks throughout the United States. The eyes were stored at 4 °C and processed within 8 h of enucleation. Primary lamina cribrosa astrocyte cultures were derived as previously described [18]. The posterior pole of the eye was dissected and the optic nerve head was freed from sclera and other neighboring tissues. The optic nerve head was sliced sagittally and under a
Results
The immunoblots in Fig. 1, Fig. 2, Fig. 3, all demonstrate that the combination of cytokines, IFNγ/IL-1β, causes a marked increase in the cellular content of NOS-2 protein in human optic nerve astrocytes in vitro. Similarly, when these astrocytes are exposed to elevated hydrostatic pressure, there are also marked increases in NOS-2 protein.
The immunoblot in Fig. 1A also shows the effects of the inhibitor of NFκB, SN-50, on the appearance of NOS-2 protein in human optic nerve astrocytes treated
Discussion
Our results confirm that several independent signal transduction pathways participate in the induction of the gene expression of NOS-2. We interpret our results to indicate that, in response to specific stimuli, downstream products of the NFκB pathway, the p38 MAP kinase pathway, and the EGFR tyrosine kinase pathway contribute to activation of the promotor region of the NOS-2 gene. We can distinguish at least one signal transduction pathway that is common for two different stimuli and two
Acknowledgements
This work was supported in part by a grant from NIH (EY 12017).
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