Journal: Nature neuroscience
Article Title: Selective inhibitors of nuclear export avert progression in preclinical models of inflammatory demyelination
doi: 10.1038/nn.3953
Figure Lengend Snippet: ( a ) Representative spinal cord sections from therapeutically treated EAE mice stained for NFH (red), SMI-32 (green), scale bar = 50 μm. ( b ) Quantification of SMI-32 signal intensity relative to NFH from. ( c ) Representative EM images from three-dimensional reconstructed blocks showing degenerating axons from vehicle-treated EAE mice, and sections from KPT-350 mice depicting intact axons with no myelin, and intact axons with split myelin lamellae, (* indicates intra-lamellar myelin debris); scale bar = 1.0 μm. ( d ) Quantification from 3D-EM data sets of axons undergoing Wallerian degeneration normalized to total axons. ( e ) Quantification from 3D-EM data sets of total axons per mm 2 . ( f ) Pie graphs representing the relative degree of cytoskeletal damage in axons from vehicle, KPT-276 or KPT-350 treated mice. Healthy axons were defined by 100% cytoskeletal arrangement. Axonal damage was then quantified based on the percentage of cytoskeletal disruption with 25% or 50% disruption as indicated by an absent, fragmented or aberrantly oriented cytoskeleton; 100% disruption when only a disrupted, or flocculent cytoskeleton were detected. At least 200 axons were counted per treatment group and at least n = 2 mice per group; scale bar = 0.1 μm. ( g ) Schematic diagram of in vivo kainic acid-induced neurotoxicity in mice. Representative confocal images of CA3 region of in vivo mouse hippocampus from mice treated with either vehicle or KPT-350 (7.5 mg/kg) for 1 hr followed by injection with kainic acid (2 mg/kg) for 18 hr. Coronal brain slices of hippocampus were stained with NFH (red), SMI-32 (green), DAPI (blue); scale bar = 100 μm. ( h ) Quantification of SMI-32 intensity. Values represent mean pixel intensity ± SEM of n=2 animals per group, from two independent experiments. ( i ) Representative IBA1 + (green) stained sections of hippocampus, sections were counterstained with DAPI (blue); scale bar = 100 μm. ( j ) Quantification of ( i ). Bar graphs represent mean ± SEM. Statistical differences in: ( b ), ( h ), and ( j ) were determined using one-way ANOVA with Tukey's correction (**p < 0.01 vs. vehicle, ***p < 0.001 vs kainic acid); ( d ), ( e ) Kruskall-Wallis tests with Dunn's correction (*p < 0.05 vs. vehicle-treated controls, n=3 animals per group).
Article Snippet: After two days, infected neurons were identified by GFP fluorescence on a live imaging microscope, and then cells were treated with 5 μM kainic acid for 2 h. Following kainic acid treatment, cells were fixed with 4% PFA, and stained with antibodies for CRM1 (1:200, Santa Cruz, cat no: sc-5595), NFH (1:400, Millipore, cat no: MAB5448), SMI-32 (1:5000, Millipore, cat no: NE1023).
Techniques: Staining, Disruption, In Vivo, Injection
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![Figure 5 RTA 408 modifies seizure progression. (A) EEG sample traces recorded for status epilepticus (SE): an asterisk indicates the beginning of status epilepticus whereas an arrow indicates termination with diazepam. (B) Typical EEG example of a spontaneous seizure post status epilepticus, expanded in the right panel. Note the rhythmical high frequency discharges at the beginning of the seizure (2) when compared to baseline (1). This rhythmic fast activity increases in amplitude (3) and decreases in frequency (4) towards the end of the seizure thus fulfilling the criteria for EEG seizure activity. (C and D) Bar charts of mean frequency (SEM) of electrographically recorded seizures following kainic acid induced status epilepticus for control rats, treated with vehicle (10% DMSO/saline) once daily for 3 days (n = 9) and RTA 408-treated rats (RTA 408 25 mg/kg once daily for 3 days; n = 9) 1–12 weeks [C; F(1,192) = 5.828, P 5 0.05] and 13–15 weeks following the first spontaneous seizure [D; F(1,18) = 10.163, P 5 0.01]; animal numbers in D: n = 4 for both groups. *P 5 0.05, **P 5 0.01 and ***P 5 0.001 by generalized log-linear mixed model followed by sequential Bonferroni post hoc test. (E) Probability distribution illustrating the probability of 0–10 seizures/day for each individual vehicle (n = 9) and RTA 408-treated rat (n = 9). (F) Bar chart summarizing mean SEM seizure probability across time; **P 5 0.01 probabilities of seizure free days were compared using Mann-Whitney U-test. (G) Seizure duration distribution for the same animals as in C, demonstrating no significant difference between vehicle or RTA408 treated animals. KA = kainic acid.](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_8645/pm29538645/pm29538645__page12_image1.jpg)
