Figure 1.
Representative diagrams used for data analysis.
A) Representative coronal striatal sections (Bregma 1.10 to −0.10 mm) obtained from the Franklin and Paxinos atlas [45]. B) Representative coronal striatal section illustrating the placement of 14 probes (236 × 140 µm area) used for the evaluation of microglial and astroglial quantification (Mercator Pro; Explora Nova, France). C) Representative coronal striatal section containing the region of interest in the dorsolateral striatum from where TH and DAT immunoreactivity was determined.
Figure 2.
Modafinil treatment prevents METH-induced decrease in TH immunoreactivity in the striatum.
Representative images of the striatum from brains of animals treated with either VEH (A, E), MOD (B, F), METH (C, G) M+M (D, H), scale bar, 500 µm, and their respective higher magnification photomicrographs (A′–H′), scale bar, 10 µm. Brain sections of animals sacrificed 48 hours (A–D) or 6 days (E–H) after the last METH injection were processed for TH immunoreactivity. Note the decrease in intensity and the changes in TH fiber morphology in the METH treated group 6 days after treatment. Average integrated density (% control) was determined in a region of interest in the dorsolateral striatum in mice (n = 4−8) sacrificed 48 hours (I) or 6 days (J) after the last METH injection. Values are expressed as mean ± SEM. Kruskal Wallis ANOVA on ranks, different letters: p<0.05.
Figure 3.
Modafinil treatment prevents METH-induced decrease in DAT immunoreactivity in the striatum.
Representative images of the striatum from brains of animals treated with either VEH (A, E), MOD (B, F), METH (C, G), or M+M (D, H), scale bar, 500 µm. Brain sections of animals sacrificed 48 hours (A–D) or 6 days (E–H) after the last METH injection were processed for DAT immunoreactivity. Note the decrease in intensity observed in the METH-treated group 6 days after treatment. Average integrated density (% control) was determined in a region of interest in the dorsolateral striatum in mice (n = 5−8 sacrificed 48 hours (I) or 6 days (J) after the last METH injection. Values are expressed as mean ± SEM. Kruskal Wallis ANOVA on ranks, different letters: p<0.05.
Figure 4.
Modafinil treatment prevents METH-induced striatal microglial activation.
Representative images of the striatum from brains of animals treated with either VEH (A, E), modafinil (MOD) (B, F), METH (C, G), or M+M (D, H), and the respective higher magnification photomicrographs (A′–H′). Brain sections of animals sacrificed 48 hours (A–D) or 6 days (E–H) after the last METH injection were labeled with ILB4 isolectin. Note the change in microglia cell morphology towards a more activated phenotype in the METH treated group 48 h after treatment, and that blood vessel were also labeled with ILB4 lectin (see arrow in panel A). Representative images of microglia showing the different morphologies observed in resting and activated states (I). Microglia averaged cell count at their resting or activated states obtained from mice (n = 4−8) sacrificed 48 hours (J) or 6 days (K) after the last METH injection. Values are expressed as mean ± SEM. One-way ANOVA followed by Tukey, different letters: p<0.001. Scale bars, 50 µm.
Table 1.
Modafinil treatment prevents METH-induced striatal amoeboid-shaped microglial differentiation.
Figure 5.
Modafinil treatment counteracts METH-induced striatal astroglial activation.
Representative images of the striatum from brains of animals treated with either VEH (A, E), MOD (B, F), METH (C, G), or M+M (D, H), scale bar, 500 µm, and their respective higher magnification photomicrographs (A′–H′), scale bar, 100 µm. Brain sections of animals sacrificed 48 hours (A–D) or 6 days (E–H) after the last METH injection were processed for GFAP immunoreactivity. Percentage of immunoreactive area was determined in mice (n = 5−8) sacrificed 48 hours (I) or 6 days (J) after the last METH injection. Values are expressed as mean ± SEM. One-way ANOVA followed by Tukey, different letters: p<0.01.
Figure 6.
Modafinil treatment prevents METH-induced increase of pro-apoptotic and decrease of anti-apoptotic markers in the striatum.
Expression of pro-apoptotic BAX (A) and anti-apoptotic Bcl2 (B) proteins in the striatum, measured by Western blot analyses. Striata were obtained 16 hours after VEH, MOD, METH or M+M treatments (n = 5−9).Values represent mean ± SEM (% of control). One-way ANOVA followed by Fisher’s LSD. different letters: p<0.05.
Figure 7.
Modafinil treatment modulates METH-induced hyperthermia.
Mice body temperatures were recorded 1 h before the first MOD injection and at different intervals thereafter. Black filled arrows indicate the time of METH injection while grey filled arrows indicate the time of MOD injection. Data is presented as mean core temperature (°C) at the indicated times ± SEM from animals treated with VEH (white circle), MOD (grey circle, dotted line), METH (Black inverted triangle), or M+M (dark grey inverted triangle, dotted line). Repeated measures two-way ANOVA followed by Fisher’s LSD, *p<0.05 vs. VEH.