Figure 4
Determination of tyrosine hydroxylase (Th) expression in the Mecp2 null mice upon different treatments. (a) Representative immunostaining of coronal brain sections from Substantia nigra pars compacta (SNpc). Midbrain dopaminergic neurons were coimmunostained for Th (green) and DAPI (4,6-diamino-2-phenolindol dihydrochloride) counterstain (blue). Scale bars, 100 μm. (b) Quantification of Th-positive staining in all aforementioned experimental groups. Bars represent the integrated density (ImageJ Fiji vs, 1.47) of the picture obtained with a SPS5 confocal microscope, × 63 objective. An increase in Th levels in Mecp2-deficient mice treated with L-Dopa and L-Dopa+Ddci is observed. (c) Representative western blot showing the levels of Th proteins in midbrain from four experimental groups. Actin was used as a loading control. (d) Dopamine concentration normalized with the respective total protein amount in caudade putamen of wild-type (WT) and Mecp2 null mice treated with vehicle or L-Dopa+Ddci (dopa-decarboxylase inhibitor). All animals were analyzed 4 weeks post treatment (8 weeks old), n=10 per group. One-way analysis of variance (ANOVA) and a post hoc test were used for statistical comparison. For the described pairwise comparisons, a Tukey HSD post hoc test was performed after the one-way ANOVA. *p<0.05, **p<0.01, and ***p<0.001.
