Fig. 1
Jacob/nsmf gene knockout does not protect against OGD- and NMDA-induced cell death. A–E Oxygen glucose deprivation (OGD) induces cell death in organotypic hippocampal slice culture irrespective of mice genotype. A Bright field and fluorescent images of propidium iodide (PI) signal in organotypic hippocampal slices from wild-type (nsmf+/+) and Jacob/nsmf constitutive knock-out animals (snmf−/−) 0, 3, 8, 12, and 24 h after 30 min OGD and control. B Scheme representing CA1, CA3, and DG areas. C–E Graphs representing the degree of PI uptake in arbitrary units (A.U.) over time (h) after OGD insult. The OGD protocol induced cell death to the same degree in CA1 (C), CA3 (D), and DG (E) irrespective of genotype. N = Control nsmf+/+: 11; OGD nsmf+/+: 13; Control nsmf−/−: 7; OGD nsmf−/−: 8 slices per group. **p < 0.01, ***p < 0.001 OGD Jac+/+ x control Jac+/+, ##p < 0.01 OGD Jac−/− x control Jac−/− by repeated measures (RM) two-way ANOVA followed by Bonferroni’s multiple comparisons test. Data represented as mean ± SEM. F–I Acute NMDA (50 µM) treatment induces cell death in OHSC irrespective of genotype. F Brightfield and PI signal in organotypic hippocampal slices from Jac+/+ Jac−/− animals after 0, 3, 8, 12, and 24 h post treatment with 50 µM NMDA or control. G, H, I Graphs representing the degree of PI uptake (A.U.) over time (h) after treatment with 50 µM NMDA. 50 µM NMDA induced cell death to the same degree in CA1, CA3, and DG irrespective of genotype. N = Control nsmf+/+: 8; NMDA nsmf+/+: 15; Control nsmf−/−: 6; NMDA nsmf−/−: 10 slices per group. **p < 0.01, ***p < 0.001, ****p < 0.0001 NMDA nsmf+/+ x control nsmf+/+ by RM two-way ANOVA followed by Bonferroni’s multiple comparisons test. Data represented as mean ± SEM. J, K MK-801 blocks NMDA-induced cell death in OHSC from C57BL6/J mice. J Brightfield and PI signal from OHSC 0, 3, 8, 12 and 24 h after treatment with 50 µM NMDA, co-application of MK-801 + 200 µM NMDA, or control. K The co-application of MK-801 completely abolished the effects of NMDA treatment. N = Control: 4; NMDA: 8; NMDA + MK-801: 4 slices per group. ***p < 0.001 vs. control by RM two-way ANOVA followed by Bonferroni’s multiple comparisons test. Data represented as mean ± SEM. Lookup table indicates the pixel intensities from 0 to 255