Figure 1.
Expression of tyrosinase in the basal ganglia of hemi-parkinsonian rats.
(A) Immunofluorescence signals using anti-tyrosinase antibody (green) and anti-GFAP antibody (red) in the mid-striatum on the control side or lesioned side of hemi-parkinsonian rats and sham-operated controls at 4 weeks after lesioning by 6-OHDA injection. (B) Double immunofluorescence staining using anti-tyrosinase antibody (green) and anti-TH antibody (red) in the substantia nigra of the intact side or lesioned side of hemi-parkinsonian rats and sham-operated controls. (A, B) Each right panel shows means integrated density of tyrosinase-IR ± SEM (n = 3). *p<0.05, **p<0.01 vs. intact side of each treated group. ##p<0.01 vs. side-matched sham-operated control group.
Figure 2.
Effects of intrastriatal transplantation of B16-F1 melanoma cells in hemi-parkinsonian rats.
(A) Rotation behavior towards contralateral side for 5 min at 5 min after injection of apomorphine (0.1 mg/kg, s.c.) was tested at 3, 7, 14 and 40 days after transplantation of B16-F1 melanoma cell (5×105 cells/4 µl PBS X2) into the right lateral striatum of hemi-parkinsonian rats or sham-operated controls. Values are means ± SEM of 3–4 rats. *p<0.05, **p<0.01, ***p<0.001 vs. each sham-PBS, ++p<0.01, +++p<0.001 vs. each 6-OHDA-PBS, ##p<0.01, ###p<0.001 vs. each pre-transplantation of B16-F1 cells. Representative photographs of hematoxylin-eosin-staining (B) and DA-IR (C) of the striatal area around the transplant at 52 days after intrastriatal transplantation of B16-F1 melanoma cells in hemi-parkinsonian rats and sham-operated controls. (C) Lower panel shows means integrated density of DA-IR ± SEM (n = 3–4). **p<0.0001 vs. control side of each treated group. ##p<0.0001 vs. side-matched PBS-injected control group.
Figure 3.
Effects of intrastriatal transplantation of tyrosinase cDNA-transfected hepatoma HLE in hemi-parkinsonian mice.
Human hepatoma HLE cells transfected with tyrosinase cDNA (HLE/tyrosinase) (1×106 cells/2 µl PBS) or empty vector (HLE) were transplanted into the right (lesioned side) lateral striatum of hemi-parkinsonian albino mice. (A) Double immunostaining of tyrosinase/L-DOPA in cultured human tyrosinase cDNA-transfected HLE cells. (B) Rotation behavior towards contralateral side over 10 min after injection of apomorphine (0.5 mg/kg, s.c.) was recorded at 3–77 days after HLE/tyrosinase transplantation. Values are means ± SEM of 4 mice. *p<0.05, **p<0.001 vs. each time-matched sham-HLE group. +p<0.05 vs. each time-matched 6-OHDA-HLE group. #p<0.01, ##p<0.001 vs. each pre-transplantation of HLE cells. (C–D) Histological changes in the striatal area around the transplant after transplantation of intrastriatal HLE/tyrosinase into hemi-parkinsonian mice. TH-immunostaining with nuclear counter-staining using methylgreen (C), tyrosinase-positive IR (D; green) and DA-positive IR (D; red) in the right lateral striatum around the transplant of hemi-parkinsonian mice at 80 days (12 weeks) after intrastriatal HLE/tyrosinase transplantation. (E) Changes in the striatal contents of L-DOPA, DA and its metabolites DOPAC and HVA of hemi-parkinsonian mice at 80 days after HLE/tyrosinase transplantation. Values are means ± SEM of 6–8 samples from 3–4 mice. *p<0.001, **p<0.0005 compared with the control side of each treated group. #p<0.05, ##p<0.005 compared with side-matched HLE cell-transplanted group.
Figure 4.
Effects of intrastriatal transplantation of primary cultured melanocytes in hemi-parkinsonian mice.
Melanocytes from albino (ICR) or black (BL) mice suspended with medium (3×105 cells/2 µl) were transplanted into the right lateral striatum of hemi-parkinsonian albino mice and sham-operated controls at 48 days after 6-OHDA lesioning. (A) Asymmetric rotation behavior towards contralateral side over 10 min after injection of apomorphine (0.5 mg/kg, s.c.) was recorded at 3–86 days after melanocyte transplantation. Values are means ± SEM of 3–4 mice. *p<0.05, **p<0.01, ***p<0.001 vs. each 6-OHDA-medium, +p<0.05, ++p<0.01 vs. each 6-OHDA-melanocytes ICR, #p<0.05, ##p<0.01 vs. each pre-transplantation of melanocytes. Histological changes in the striatal area around the transplant after intrastriatal melanocyte transplantation into hemi-parkinsonian mice. Hematoxylin-eosin-staining (B), tyrosinase-positive IR (C), and DA-positive IR (D) in the right lateral striatum around the transplant of hemi-parkinsonian albino mice and sham-operated controls at 88 days (12 weeks) after intrastriatal transplantation of melanocytes obtained from albino (ICR) or black (C57BL) mice.
Figure 5.
Tyrosinase-positive IR (A) and DA-IR (B) in the right lateral striatum around the transplant of hemi-parkinsonian albino mice and sham-operated controls at 88 days after intrastriatal transplantation of melanocytes obtained from albino (ICR) or black (C57BL) mice.
Values are means integrated density of tyrosinase-IR or DA-IR ± SEM (n = 3–4). *p<0.005, **p<0.0001 vs. intact control side of each treated group. #p<0.005, ##p<0.0001 vs. side-matched medium-injected control group. $p<0.005, $$p<0.0001 vs. side-matched melanocytes from ICR mice-transplanted group. +p<0.05, ++p<0.0005 vs. side- and treatment-matched sham-operated.