Experiment 3- Striatal LID markers expression after administration of doxycycline
In this experiment, the immunoreactivity of FOS-B (a canonical protein associated with LID), proteins typical of neuroinflammation such as COX-2, GFAP, OX-42, MMPs, and the presence of ROS were evaluated. The experimental groups comprised parkinsonian rats with already established LID that were receiving acute doxycycline treatment and parkinsonian rats receiving doxycycline treatment since day one as well as the respective controls. An individual experimental group assembly was prepared for each analysis described below.
Experiment 3.1. Striatal LID markers expression in rats acutely treated with doxycycline once dyskinesia was present
(i) Immunohistochemical reaction for COX-2, GFAP, OX-42 in the experimental groups: 6-OHDA+vehicle+L-DOPA, (n=7); 6-OHDA+doxycycline+L-DOPA (n=8) and the respective controls [no-lesion+vehicle+vehicle; (n=6), 6-OHDA+vehicle+vehicle (n=5), 6-OHDA+doxycycline+vehicle (n=5)].
(ii) MMP-3 immunoblotting, MMP-2 and MMP-9 gel zymography in the striatum of the experimental groups: 6-OHDA+vehicle+L-DOPA (n=7), 6-OHDA+doxycycline+L-DOPA (n=6), and the respective controls [no-lesion+vehicle+vehicle (n=6), 6-OHDA+vehicle+vehicle, (n=6), 6-OHDA+DOX+vehicle (n=6)].
(iii) MMP-2/-9 in situ gelatinolytic zymography and ROS histochemistry analysis in the brain sections of the experimental groups: 6-OHDA+vehicle+L-DOPA (n=7), 6-OHDA+doxycycline+L-DOPA (n=7), and the respective controls [no-lesion+vehicle+vehicle (n=6), 6-OHDA+vehicle+vehicle (n=6), 6-OHDA+doxycycline+vehicle (n=6)].