Figure 1

NOS2 expression during glaucoma progression. (A) Optic nerve head Nos2 RNA expression. Relative expression levels of Nos2 at different stages of glaucoma were determined using ΔCT ratios (see Methods). 'Control' eyes from 4 months old DBA/2J mice were considered to be pre-glaucomatous and showed no axon damage. 'Sample' eyes from 10.5 months old DBA/2J mice were assessed for glaucomatous axon damage and grouped into the no or early, moderate or severe stages of damage (see Methods). Values given are relative to pre-glaucoma controls. Nos2 expression was variable within each disease group (indicated by the SEM bars). Overall it decreased comparing pre-glaucoma with no or early glaucoma and increased modestly in eyes with moderate glaucoma (p < 0.05). (B) NOS2 protein localization in LPS-treated kidney (above) and no primary control (below). The NOS2 antibody shows strong and specific NOS2 staining (Red) in LPS-treated kidneys, with no non-specific binding of the secondary antibody. (C) NOS2 protein localization in the optic nerve heads of 10.5 months old DBA/2J mice, with the lower panels showing higher magnification of the boxed regions. There was no obvious correlation between NOS2 localization and glaucoma progression, although rare NOS2 positive cells were seen in eyes with severe glaucoma (arrow). All sections were processed, stained and imaged under identical conditions. The images shown here are representative of three different sections from three different eyes with the three different stages of glaucoma. Bars = 75 μm.