Article Figures & Data
Figures
- Fig. 1.
Caspase-1 and caspase-3 mRNA levels are altered in the spinal cords of transgenic mSOD1 mice. A, B, Trend toward increased levels of caspase-1 mRNA in transgenic mSOD1 mice at the beginning of symptoms and at end-stage. A, C, Significant increased caspase-3 mRNA levels in transgenic mSOD1 mice at the beginning of symptoms and at end-stage; *p < 0.05 higher than age-matched nontransgenic controls, Newman–Keulspost hoc test. N, Nontransgenic;A, asymptomatic; O, onset of symptoms;E, end-stage.
- Fig. 2.
Activation of caspase-1 and caspase-3 in the spinal cords of transgenic mSOD1 mice. A, B, By 12 weeks of age the levels of pro-caspase-1 (45 kDa; black bars) decrease, whereas those of its cleaved fragment (20 kDa; white bars) increase in transgenic mSOD1 mice. A, C, A few weeks later the same phenomenon occurs for pro-caspase-3 (32 kDa;black bars) and its cleaved fragment (17 kDa;white bars); *p < 0.05 and †p < 0.01 different from age-matched nontransgenic controls, Newman–Keuls post hoc test.N, Nontransgenic; A, asymptomatic (at 4, 6, 8, 12 weeks of age); O, onset of symptoms;E, end-stage; C, cerebellum.
- Fig. 3.
Increased caspase-1- and caspase-3-like activities in the spinal cords of transgenic mSOD1 mice. Caspase-1-like activity (black bars) increases before caspase-3-like activity (white bars), but both peak at the beginning of symptoms. Data are mean ± SEM for 5–15 mice per group; *p < 0.05 and **p < 0.01 higher than age-matched nontransgenic controls, Newman–Keulspost hoc test. N, Nontransgenic;A, asymptomatic (at 4, 6, 8, 12 weeks of age);O, onset of symptoms; E, end-stage;C, cerebellum.
- Fig. 4.
In nontransgenic controls a large number of cells from the lumbar segment are strongly immunoreactive for pro-caspase-1 (A) and pro-caspase-3 (D); those cells have a neuronal morphology (see higher magnification inB and E). In end-stage transgenic mSOD1, there is a dramatic loss of pro-caspase-1-positive (C) and pro-caspase-3-positive (F) neurons in the lumbar segment. Conversely, specific immunostaining for CM1 (G; seearrow) and fractin (H; seearrow) is seen only in symptomatic transgenic mSOD1 mice within apoptotic cells (see insets;arrowheads indicate apoptotic chromatin clumps) and colocalizes with the neuronal marker neurofilament (I), but not with the glial marker GFAP (J). Scale bars: A, C, D, F–H, 100 μm; B, E, I, J, 20 μm.
- Fig. 5.
Bcl-2 delays caspase activation. Western blot analyses (A–D) show that transgenic mSOD1/Bcl-2 mice that are aged-matched (AM) with end-stage (E) transgenic mSOD1 exhibit significantly lower levels of cleaved caspase-1 (A, C) and caspase-3 (B, D) than end-stage transgenic mSOD1. However, end-stage transgenic mSOD1/Bcl-2 mice exhibit levels of cleaved caspase-1 (A, C) and caspase-3 (B, D) comparable with end-stage transgenic mSOD1. A similar situation is found for the activity of caspase-1 and caspase-3 (E); *p < 0.05 higher and †p < 0.05 lower than end-stage transgenic mSOD1 and mSOD1/Bcl-2 mice, Newman–Keuls post hoc test.
Tables
mSOD1 Nontransgenic mSOD1/Bcl-2 Beginning of symptoms End-stage Age-matched1-a Age-matched1-a End-stage MN 65 ± 3* 32 ± 21-160 74 ± 2 61 ± 2* 34 ± 21-160 CM1 × 10 3.3 ± 0.6 2.7 ± 0.6 0 1.2 ± 0.5† 3.1 ± 0.9 Fractin × 10 3.7 ± 0.6 2.8 ± 0.8 0 1.5 ± 0.4† 2.7 ± 0.5 ↵F1-a Killed simultaneously with transgenic mSOD1 end-stage littermates. Cells were counted in the lumbar segment of the spinal cord. Data represent mean per section ± SEM for three to five mice per group. MN, Motor neuron.
↵* p < 0.05 and
↵F1-160 p< 0.01 lower than wild-type mice;
↵† p < 0.05 lower than beginning of symptoms and end-stage transgenicmSOD1, Newman–Keuls post hoc test.
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