Journal: Brain Communications

Article Title: α-Amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor and RNA processing gene dysregulation are early determinants of selective motor neuron vulnerability in a mouse model of amyotrophic lateral sclerosis

doi: 10.1093/braincomms/fcac081

Figure Lengend Snippet: Validation of differentially expressed genes using qRT-PCR. ( A–E ) qPCR verification of the expression of genes involved in biological process identified as enriched by GO analysis compared with WT control. (F) Fold change expression of Ca 2+ -permeable AMPAR subunit Gria1 , Gria3 and Gria4 mRNAs, relative to WT motor neurons at E12.5. (G) Relative expression of Adarb1 mRNA in SOD1 G93A motor neurons at E12.5. ( H ) Schema showing the position of the fully complementary miR-124 target site at the 5′-end of the mouse Gria2 , 3′-UTR. The seed region of miR-124 is shown. Data represent mean ± SEM, unpaired student t -test, n = 5–7 biological replicates, * P < 0.05.

Article Snippet: Primary antibodies were as follows: chicken anti-GFP (1:1000; Abcam; AB13970), rabbit anti-NeuN (1:1000; Abcam; AB104225), goat anti-ChAT (1:500; Abcam; AB34419) and guinea pig anti-AMPA receptor 2 subunit (GluA2) (1:500; Alomone Labs; AGP-073).

Techniques: Quantitative RT-PCR, Expressing

Journal: Brain Communications

Article Title: α-Amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor and RNA processing gene dysregulation are early determinants of selective motor neuron vulnerability in a mouse model of amyotrophic lateral sclerosis

doi: 10.1093/braincomms/fcac081

Figure Lengend Snippet: Expression of GluA2 in spinal cords of embryonic SOD1 G93A mice. Cross-sections of lumbar spinal cord from WT (HB9:GFP; WT) and SOD1 G93A (SOD1 G93A ; HB9:GFP) mice at (A–J) E12.5 and ( K–T ) E17.5. Double-immunolabelling for GFP, GluA2 and NeuN (Neuronal nuclei). Plots represent quantification analysis of GluA2 signal intensity in HB9:GFP motor neurons at ( U ) E12.5 and ( V ) E17.5. Data represent mean ± SEM, unpaired student t -test performed on n = 4 biological replicates, ∼50 neurons analysed per biological replicate, * P < 0.05. Scale bars 50 μm.

Article Snippet: Primary antibodies were as follows: chicken anti-GFP (1:1000; Abcam; AB13970), rabbit anti-NeuN (1:1000; Abcam; AB104225), goat anti-ChAT (1:500; Abcam; AB34419) and guinea pig anti-AMPA receptor 2 subunit (GluA2) (1:500; Alomone Labs; AGP-073).

Techniques: Expressing

Journal: Brain Communications

Article Title: α-Amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor and RNA processing gene dysregulation are early determinants of selective motor neuron vulnerability in a mouse model of amyotrophic lateral sclerosis

doi: 10.1093/braincomms/fcac081

Figure Lengend Snippet: Expression of GRIA2 and ADAR2 in iPSC motor neurons derived from ALS patients with SOD1 mutations and healthy control lines. Representative images of iPSC mature motor neurons derived from ( A–E ) healthy control line and ( F–J ) SOD1 I114T line, immunolabelled with ChAT, GluA2 and TUJ1, counterstained with Hoechst. ( K ) Plot represents quantification analysis of GluA2 signal intensity in iPSC motor neurons. Data represent mean ± SEM, unpaired student t -test performed on n = 3 biological replicates, 50 neurons analysed per biological replicate. (L) Fold change expression of GRIA2 in SOD1 lines, compared with healthy control line determined by qRT-PCR. ( M ) Fold change expression of ADAR2 in SOD1 lines, compared with healthy control line determined by qRT-PCR. Data represent mean ± SEM, n = 3 biological replicates, one-way ANOVA with Dunnett's multiple comparison test, * P < 0.01, ** P < 0.005. Scale bars 50 μm.

Article Snippet: Primary antibodies were as follows: chicken anti-GFP (1:1000; Abcam; AB13970), rabbit anti-NeuN (1:1000; Abcam; AB104225), goat anti-ChAT (1:500; Abcam; AB34419) and guinea pig anti-AMPA receptor 2 subunit (GluA2) (1:500; Alomone Labs; AGP-073).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR

Journal: Frontiers in Neuroscience

Article Title: Perampanel Add-on to Standard Radiochemotherapy in vivo Promotes Neuroprotection in a Rodent F98 Glioma Model

doi: 10.3389/fnins.2020.598266

Figure Lengend Snippet: Synergistic impact of PER and RCT on glutamatergic network activity and GluA2 expression in acute F98 glioma slices. (A) One week after stereotactic injection of 1 × 10 5 F98 cells, RCT with fractionated irradiation (5 × 4 Gy), concurrent administration of temozolomide (5 × 30 mg/kg bw) and an anticonvulsive treatment with PER (15 mg/kg bw/day) was started. After a total of 14–15 days animals were sacrificed and brain slices were prepared for electrophysiological analysis. The slices were exposed to aCSF 0 mM Mg 2+ and 5 μM gabazine and field potential recordings were performed in surrounding tissue; n = 17–48 number of measurements (brain slices were from a total of 35 rats); * p < 0.05 (Kruskal–Wallis test followed by post hoc analysis (Dunn’s test); # < 0.05 versus contralateral equivalent ( U test). (B) AMPA receptor subunit GluA2 expression was determined in the tumor area and peritumoral tissue of the ipsilateral hemisphere. Glu2A (red) immunofluorescence was normalized to peritumoral immunofluorescence intensity of the vehicle controls without PER or RCT (see section “Materials and Methods”). Nuclei were counterstained with DAPI (blue); white dotted line represents arithmetic mean, black solid line represents median. Significant difference between peritumoral and glioma fluorescence for each group was determined ( n = 7–8 animals per group; * p < 0.05; U test).

Article Snippet: Additionally, the expression of the AMPA receptor subunit GluA2 in the tumor area, the peritumoral tissue and the contralateral hemisphere was determined using anti-GluA2 antibody (Alomone labs; AGC-005; Jerusalem, Israel) as the primary antibody and anti-rabbit Cyanine5 as the secondary antibody.

Techniques: Activity Assay, Expressing, Injection, Irradiation, Immunofluorescence, Fluorescence