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Fig. 1. Stereotaxic 6-OHDA injection leads to loss of DA cells in the
diencephalon and the mesencephalon. (A,B) Low- and high-power
photomicrographs of TH+ cells in the tegmental area of the newt
midbrain. (C) TH+ fibres in the striatum from a
sham-lesioned animal. (D) High-power photomicrograph of the boxed area
in C. (E,F) TH+ cells in the TM and (G)
TH+ fibres in the striatum from a 97% lesioned animal 3 days after
6-OHDA injection. (H) High-power photomicrograph of the boxed area in
G. (I) A significant loss of TH cells was seen within the ventral
diencephalon-mesencephalon 3 days post-lesioning (n=16 lesioned,
n=14 sham, n=7 control; mean±s.d.; Student's
t-test; ***, P<0.001). (J)
Colocalisation of TUNEL+ and TH+ cells in the TM 3 days
after lesioning. (K) Significantly more TUNEL+ cells and
numerous TUNEL+ TH+ cells were observed in lesioned as
compared with sham-lesioned animals. No double-positive cells were seen in
sham-lesioned animals (mean±s.d.; Student's t-test;
***, P<0.001). (L') Schematic
representation of the lesioning apparatus. (L'') The location of
the burr hole and injection site (*), at the junction of the
frontal and parietal bones. (M) A significantly higher percentage of
pyknotic cells (ANOVA with Tukey post-hoc test; P<0.01) was seen
12 hours (5±1.2; n=7) and 24 hours (11.9±2.9;
n=7) after lesioning as compared with shams (12 and 24 hour
sham-lesioned animals pooled; 1.7±1.6; n=10). Note the
increase in the percentage of pyknotic cells with increasing lesion size, and
the increase in the percentage of pyknotic cells in the lesioned animals
between 12 and 24 hours. (N,O) 6-OHDA injection leads to a significant
loss of cells (DAPI), including neurons (NeuN), within the TM, and more
specifically to the loss of DA markers [Nurr1, engrailed 1 (En1), TH].
n=3-7; mean±s.e.m.; Student's t-test; ***,
P<0.001. Scale bars: 1 mm in A,C,E,G; 150 µm in B,F,J; 250
µm in D,H.
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