Journal: The American Journal of Pathology
Article Title: The Canonical Wnt Signaling Pathway Inhibits the Glucocorticoid Receptor Signaling Pathway in the Trabecular Meshwork
doi: 10.1016/j.ajpath.2021.02.018
Figure Lengend Snippet: Wnt3a reduces ocular hypertension in dexamethasone (Dex)–treated mice. A: GTM3 cells were transfected with the glucocorticoid receptor (GR) signaling reporter vector and transduced with the Ad5-CMV-GFP or Ad5-CMV-Wnt3a+mCherry before luciferase assays. Data were analyzed using one-way analysis of variance and Sidak post-hoc tests. B: Three-month–old female C57BL/6J mice received binocular periocular Dex-acetate (Dex-Ac) injection once a week throughout the study (arrow on the left) after baseline intraocular pressure (IOP) establishment. After ocular hypertension development, Ad5-CMV-GFP was injected intravitreally into one eye and Ad5-CMV-Wnt3a+mCherry was injected into the fellow eye (arrow on the right). Mouse IOP was measured under isoflurane anesthesia about once a week throughout the study in a masked manner. Paired t-test was used to compare IOP at each time point. C: Hematoxylin and eosin (H&E) staining of a mouse eye section transduced with Ad5-Wnt3a-mCherry. D: Immunostaining of a section adjacent to C using the anti-mCherry antibody (red; pseudocolor). Data are given as means ± SD of GR signaling activity (A) or means ± SEM of IOP (B). N = 8 (A); N = 17 (B). ∗P < 0.05; ∗∗P < 0.01, and ∗∗∗∗P < 0.0001. Scale bar = 40 μm (C).
Article Snippet: After ocular hypertension development (about 2 weeks after initial Dex-acetate injection), 3 × 10 7 plaque-forming units Ad5-CMV-GFP were intravitreally injected into the right eye, and 3 × 10 7 plaque-forming units Ad5-CMV-mouseWnt3a+mCherry (co-expression, not a fusion protein) (Vector Biolabs, Malvern, PA) were intravitreally injected into the left eye under anesthesia.
Techniques: Transfection, Plasmid Preparation, Transduction, Luciferase, Injection, Staining, Immunostaining, Activity Assay
