SUPPLEMENTARY FIGURE LEGEND
Suppl. Figure 1. Photomicrography of coronal brain sections illustrating the loss of striatum TH positive fibers (A–C; A’-C) and substantia nigra compacta TH positive neurons (D-F; D’-F); (A’, B’, C’, D’, E’, F’) represents the lesion side. Scale bars=1200 µm.
Suppl. Figure 2. Doxycycline or COL-3 single administration to L-DOPA primed rats or co-administration of L-DOPA and doxycycline reduces individual AIMs development. (A, B, C) show doxycycline single administration effects on axial (p=0.069), limb (p < 0.01) and orofacial (p < 0.05) AIMs, respectively over 60 min on day 15 compared to day 14 LID measurement (paired t-test; the points represent data from individual animals). (D, E, F) show the co-administration of L-DOPA and doxycycline effects on the axial, limb, and orofacial AIMs, respectively over 60 min (p<0.0001, unpaired t-test, data as box and whiskers plus minimum and maximum values, the point values represent data from individual animals). (G, H, I) show COL-3 single intraventricular administration effects on axial (p < 0.01), limb (p < 0.01) and orofacial (p < 0.05) AIMs, respectively over 60 min on day 15 compared to day 14 (paired t-test; the points represent data from individual animals). #compared to 6-OHDA+vehicle+L-DOPA; +compared to 6-OHDA+vehicle+L-DOPA. AIMs: abnormal involuntary movements; DOX: doxycycline; L-DOPA, L-3,4-dihydroxyphenylalanine.
Suppl. Figure 3. Morphological quantification of the astrocytes and microglia reactivity. Doxycycline reduces activation of the astrocytes (GFAP, glial fibrillary acid protein labeling astrocytes) and microglia (OX-42 CD11b/c-beta-integrin marker of microglia) associated with LID. The analysis was performed in the striatum of 6-OHDA-lesioned rats expressing LID, without and with doxycycline treatment. Animals were sacrificed 1 h after the last L-DOPA injection. Morphological quantification of the GFAP and OX42 reactivity is shown in the bar charts and represents the average process number (A) (D),average process length (B) (E), and process intersection number (C) (F), respectively. A two-way ANOVA followed by the Bonferroni test was used to check for statistical significance: *compared to 6-OHDA+vehicle+vehicle; #compared to 6-OHDA+vehicle+L-DOPA.
Suppl. Figure 4. Substrate gel zymography quantification of striatal MMP-2 and MMP-9 isoforms, following acute and chronic doxycycline administration in parkinsonian rats (A-B) : 6-OHDA-lesion and L-DOPA treatment increased MMP-2 and MMP-9 in the striatum. A single dose of doxycycline reduced MMP-9 in the striatum of lesioned rats, and L-DOPA treated rats. (C-D): The administration of L-DOPA increased the presence of MMP-2 and MMP-9 in the lesioned striatum. The co-administration of doxycycline during L-DOPA (or its vehicle) treatment reduced both MMP-2 and MMP-9 in the striatum of lesioned rats, and L-DOPA treated rats. A one-way ANOVA followed by a Bonferroni test was used to check for statistical significance (p<0.05). *compared to 6-OHDA+vehicle+vehicle; #compared to 6-OHDA+vehicle+L-DOPA. AIMs abnormal involuntary movements; DOX, doxycycline; L-DOPA, L-3,4-dihydroxyphenylalanine; MMPs, matrix metalloproteinase