Journal: Cell Death and Differentiation
Article Title: Acidotoxicity and acid-sensing ion channels contribute to motoneuron degeneration
doi: 10.1038/cdd.2012.158
Figure Lengend Snippet: Expression of asic1, asic2 and asic3 during ALS disease progression. (a) Co-staining of lumbar spinal cords from tg SOD1- and asic1a-deficient mice with anti-ASIC1/ASIC2 (red) and anti-NeuN/GFAP (green) antibodies. Arrowheads show co-localization of ASIC1 and ASIC2 with NeuN on cell soma of neurons with motoneuron morphology. Scale bar=20 μm. (b) asic1, −2 and −3 mRNA expression in the transgenic (tg) SOD1 mouse; lumbar spinal cord lysates show a significant upregulation of the asic1a and asic2a mRNA; *P<0.01. Data displayed as mean±S.D.; n=6/group. (c) Co-staining of lumbar spinal cords across disease progression in the tg SOD1 mouse with anti-ASIC1/ASIC2 (red) and anti-NeuN/smi-32 (green) antibodies. Co-localization of ASIC antibodies with motoneurons is indicated by arrowheads. Scale bar=15 μm. (d) An increase in ASIC1 and ASIC2 protein levels in the tg SOD1 mouse; *P<0.01. Data displayed as mean±S.D.; n=6/group. (e) A representative image of ASIC1 and ASIC2 protein (double band) levels by western blotting, n=3 experiments with tubulin as loading control. (f) ASIC2 immunohistochemistry in ventral motoneurons of lumbar spinal cord sections from ALS and non-ALS patients. n=3/group. Scale bar=50 μm
Article Snippet: Sections were incubated with a polyclonal ASIC2 antibody (1 : 200, Alomone Labs, Jerusalem, Israel), biotinylated secondary antibody (1 : 500; Jackson Laboratories) and visualized using Sigma fast 3,3-diaminobenzadine tablets.
Techniques: Expressing, Staining, Transgenic Assay, Western Blot, Immunohistochemistry