Journal: Molecular psychiatry

Article Title: NitroSynapsin ameliorates hypersynchronous neural network activity in Alzheimer hiPSC models

doi: 10.1038/s41380-020-0776-7

Figure Lengend Snippet: a Spontaneous action potentials (sAP) at resting membrane potential (RMP). WT/WT hiPSC-derived cerebrocortical neuron data in black, M146V/WT and APP swe /WT in red. b Quantification of sAP frequency. c Representative traces of glutamate-evoked currents from neurons voltage-clamped at −70 mV. d Glutamate-evoked current density. e , f Inhibition by 5–10 μM memantine or NitroSynapsin. Representative traces of WT/WT, M146V/WT and APP swe /WT neurons clamped at −70 mV (e). Percent inhibition by memantine (Mem) or NitroSynapsin (NitroSyn) in M146V/WT and APP swe /WT neurons vs. their respective WT/WT isogenic controls (f). Sample size represents total number of neurons recorded and is listed above bars. Data are mean ± SEM. Exact p values are listed above bars in this and subsequent figures unless otherwise specified. Statistical significance analyzed by ANOVA with post hoc Dunnett’s test.

Article Snippet: Recordings were performed in BrainPhys medium at 37 °C using the ‘neural spikes analog mode’ setting with a sampling frequency of 12.5 kHz at baseline (prior to) and after 5 μM NitroSynapsin treatment (Axion Biosystems Maestro Axis Software version 2.4.2) on a Maestro MEA (Axion Biosystems).

Techniques: Derivative Assay, Inhibition

Journal: Molecular psychiatry

Article Title: NitroSynapsin ameliorates hypersynchronous neural network activity in Alzheimer hiPSC models

doi: 10.1038/s41380-020-0776-7

Figure Lengend Snippet: WT/WT hiPSC-derived cerebrocortical neuron data in black, M146V/WT and APP swe /WT in red. a Representative calcium imaging traces showing decrease in [Ca 2+ ] i and transients after application of 5 μM NitroSynapsin. Dotted line represents decrease in [Ca 2+ ] i . b Quantification of difference in normalized fluorescence (ΔF/F 0Drug - ΔF/F 0Control ) in response to 5 μM memantine (Mem) or equimolar NitroSynapsin (NitroSyn). c Quantification of calcium event frequency before and after application of 5 μM NitroSynapsin. d Fluo-4 images of WT/WT and ΔE9/WT cultures showing [Ca 2+ ] i before (control) and after application of NitroSynapsin (5 μM NitroSyn). Scale bar: 10 μm. e Quantification of difference in normalized fluorescence (ΔF/F 0Drug - ΔF/F 0Control ) in response to 5 μM memantine (Mem) or equimolar NitroSynapsin (NitroSyn). f Noise-floor normalized power spectrum showing concentration of power in low frequencies (<1 Hz) and at 6 Hz (theta oscillation). WT in black; AD in red. g Quantification of mean log power (0.2–1 Hz) in WT and AD cultures. h Quantification of mean log power difference (0.2–1 Hz) after treatment with 5 μM NitroSynapsin in WT and AD cultures (post - pre). Data are mean ± SEM. Statistical significance analyzed by two-tailed unpaired Student’s t-test (*) in b, e, g and h; by ANOVA with post hoc Dunnett’s test (*) for multiple comparisons to WT; and by Sidak’s test (#) corrected for multiple comparisons between selected pairs in c.

Article Snippet: Recordings were performed in BrainPhys medium at 37 °C using the ‘neural spikes analog mode’ setting with a sampling frequency of 12.5 kHz at baseline (prior to) and after 5 μM NitroSynapsin treatment (Axion Biosystems Maestro Axis Software version 2.4.2) on a Maestro MEA (Axion Biosystems).

Techniques: Derivative Assay, Imaging, Fluorescence, Concentration Assay, Two Tailed Test

Journal: Molecular psychiatry

Article Title: NitroSynapsin ameliorates hypersynchronous neural network activity in Alzheimer hiPSC models

doi: 10.1038/s41380-020-0776-7

Figure Lengend Snippet: a-c Representative heat maps and raster plots of MEA recordings in WT/WT (a), M146V/WT (b), and APP swe /WT (c) cultures before ( left ) and after ( center and right ) treatment with 5 and 10 μM NitroSynapsin, respectively. Red boxes represent examples of network bursts. d-f Quantification of MEA recordings. Mean firing rate (d). Network burst frequency quantification (e). Quantification of synchrony index (f). Data are mean ± SEM. Sample size listed above bar graphs (*, # p < 0.05; **, ## p < 0.01; ***, ### p < 0.001; #### p < 0.0001 by ANOVA with post-hoc Dunnett’s test (*) for multiple comparisons to WT and Sidak’s test ( # ) corrected for multiple comparisons between selected pairs).

Article Snippet: Recordings were performed in BrainPhys medium at 37 °C using the ‘neural spikes analog mode’ setting with a sampling frequency of 12.5 kHz at baseline (prior to) and after 5 μM NitroSynapsin treatment (Axion Biosystems Maestro Axis Software version 2.4.2) on a Maestro MEA (Axion Biosystems).

Techniques:

Journal: Molecular psychiatry

Article Title: NitroSynapsin ameliorates hypersynchronous neural network activity in Alzheimer hiPSC models

doi: 10.1038/s41380-020-0776-7

Figure Lengend Snippet: a Representative images of 2-month old WT/WT, M146V/WT, and APP swe /WT organoids stained for Nestin, TBR2, and CTIP2 showing cortical layers. b Representative raster plots of MEA recordings in 3-month old WT/WT, M146V/WT, and APP swe /WT organoids before (left) and after treatment with 10 μM memantine (center) or 10 μM NitroSynapsin (right). Red boxes represent examples of network bursts. c - f Quantification of MEA recordings. Mean firing rate; percentage inhibition with 10 μM memantine or NitroSynapsin; network burst frequency, and synchrony index. Data are mean ± SEM. Sample size listed in the bar graphs (*, # p < 0.05; **, ## p < 0.01 by ANOVA with post-hoc Dunnett’s test (*) for multiple comparisons to WT and Sidak’s test ( # ) corrected for multiple comparisons between selected pairs).

Article Snippet: Recordings were performed in BrainPhys medium at 37 °C using the ‘neural spikes analog mode’ setting with a sampling frequency of 12.5 kHz at baseline (prior to) and after 5 μM NitroSynapsin treatment (Axion Biosystems Maestro Axis Software version 2.4.2) on a Maestro MEA (Axion Biosystems).

Techniques: Staining, Inhibition