af-dx 116  (Bio-Techne corporation)

Journal: bioRxiv

Article Title: Pathogenic MTOR somatic variant causing focal cortical dysplasia drives hyperexcitability via overactivation of neuronal GluN2C NMDA receptors

doi: 10.1101/2023.12.01.569539

Figure Lengend Snippet: (A) Representative traces of averaged sEPSCs recorded in MTOR-WT (left) and MTOR-p.S2215F (right) spiny stellate neurons in: black curve, drug-free (No drug) condition, representing total component of sEPSC; red curve, after adding GluN2C-mediated NMDAR blocker DQP 1105 (10 µM), representing the non-GluN2C-mediated component of sEPSC; and blue curve, after subtraction of the non-GluN2C-mediated component from total sEPSC, hence representing GluN2C-mediated component of sEPSC (B,C) Charge transfers were calculated in (B) MTOR-WT neurons and in (C) mutant MTOR-p.S2215F neurons, as areas between peak and 200 ms after the peak of the total sEPSC: with No drug; with GluN2C blocker DQP1105 (DQP; 10 µM); with DQP1105 (DQP; 10 µM) + GluN2B blocker Ro25-6981 (DQP+Ro; 1 µM); and with total NMDAR blocker APV (50 µM). **: p<0.01; *: p<0.05; ns: not significant. Mixed effect analysis with multiple comparison test.

Article Snippet: All recordings were done in ACSF-2 without or with 10 μM DQP 1105 (Tocris Bio-Techne, France), 10 μM DQP 1105 + 1μM Ro 25-6981 (Tocris Bio-Techne, France), or 50 μM APV (Hello Bio, UK) at room temerature (22-25°C).

Techniques: Mutagenesis, Comparison

Journal: bioRxiv

Article Title: Pathogenic MTOR somatic variant causing focal cortical dysplasia drives hyperexcitability via overactivation of neuronal GluN2C NMDA receptors

doi: 10.1101/2023.12.01.569539

Figure Lengend Snippet: (A) Representative traces of spontaneous activity recorded from green fluorescent electroporated areas in Mg 2+ containing ACSF medium from (top trace) mutant MTOR-p.S2215F slice and (bottom trace) MTOR-WT slice (B) Summary data for spiking frequency. MTOR-WT, n = 8 slices from 7 pups; MTOR-p.S2215F, n = 10 slices from 9 pups. *: p<0.05. Mann Whitney test, two-tailed (C) Representative traces of spontaneous activity recorded in mutant MTOR-p.S2215F slices (n = 10 from 9 pups) obtained in Mg 2+ containing ACSF with: (top) no drug; (middle) GluN2C blocker DQP1105 (DQP, 10 μM); and (bottom) total NMDAR blocker APV (50 μM) (D) Summary data for spiking frequency. **: p<0.01; *: p<0.05. Friedman test with Dunn’s multiple comparison test (E) Representative traces of spontaneous activity recorded from the electroporated areas in Mg 2+ containing ACSF medium from: (top trace) slice from vehicle-treated, mutant MTOR-p.S2215F pup; and (bottom trace) slice from rapamycin-treated, mutant MTOR-p.S2215F pup without (No drug, left) or with DQP 1105 (10 µM, right) (F) Summary data for spiking frequency; n = 5 slices from 2 pups in each condition. *: p < 0,05. Mann-Whitney test, two-tailed (G) Summary data for spiking frequency without or with DQP 1105 for MTOR-p.S2215F + vehicle (left, n = 5 slices from 2 pups) and MTOR-p.S2215F + Rapamycine (right, n = 5 slices from 2 pups). Note the statistical trend towards significant effect of DQP 1105 in the MTOR-p.S2215F + vehicle condition, as expected. Wilcoxon test, two-tailed.

Article Snippet: All recordings were done in ACSF-2 without or with 10 μM DQP 1105 (Tocris Bio-Techne, France), 10 μM DQP 1105 + 1μM Ro 25-6981 (Tocris Bio-Techne, France), or 50 μM APV (Hello Bio, UK) at room temerature (22-25°C).

Techniques: Activity Assay, Mutagenesis, MANN-WHITNEY, Two Tailed Test, Comparison

Journal: bioRxiv

Article Title: Pathogenic MTOR somatic variant causing focal cortical dysplasia drives hyperexcitability via overactivation of neuronal GluN2C NMDA receptors

doi: 10.1101/2023.12.01.569539

Figure Lengend Snippet: (A) Representative traces of spontaneous activity recorded from electroporated areas in Mg2+ containing ACSF medium from (top trace) MTOR-WT slice and (bottom trace) MTOR-p.S2215F slice from P9 pups, without (No drug, left) or with DQP 1105 (10 µM, right) (B) Summary data for spiking frequency. MTOR-WT, n = 5 slices from 4 pups; MTOR-p.S2215F, n = 7 from 6 pups. ns: not significant; Mann-Whitney test, two-tailed (C) Summary data for spiking frequency without or with DQP 1105 for MTOR-WT (left) and MTOR-p.S2215F (right). Data shown are values normalized to the No drug condition. *: p<0.05; ns: not significant; Wilcoxon test, two-tailed (D) Representative traces of spontaneous activity recorded from electroporated areas in Mg2+ containing ACSF medium from (top trace) MTOR-WT slice and (bottom trace) MTOR-p.S2215F slice from P20-21 pups without (No drug, left) or with DQP 1105 (10 µM, right) (E) Summary data for spiking frequency. MTOR-WT, n = 7 slices from 4 pups; MTOR-p.S2215F, n = 8 slices from 5 pups. ns: not significant. Mann Whitney test, two-tailed (F) Summary data for spiking frequency without or with DQP 1105 for MTOR-WT (left) and MTOR-p.S2215F (right). Data shown are values normalized to the No drug condition. **: p<0.01; ns: not significant. Wilcoxon test, two-tailed.

Article Snippet: All recordings were done in ACSF-2 without or with 10 μM DQP 1105 (Tocris Bio-Techne, France), 10 μM DQP 1105 + 1μM Ro 25-6981 (Tocris Bio-Techne, France), or 50 μM APV (Hello Bio, UK) at room temerature (22-25°C).

Techniques: Activity Assay, MANN-WHITNEY, Two Tailed Test