Journal: Current Research in Neurobiology

Article Title: Gene delivery to neurons in the auditory brainstem of barn owls using standard recombinant adeno-associated virus vectors

doi: 10.1016/j.crneur.2020.100001

Figure Lengend Snippet: Examples for the expression of serotypes rAAV2/1 and rAAV2/5 in nucleus magnocellularis . A–D: Expression of GFP, indicating rAAV2/1, in NM cell bodies and fibers, three weeks after injection (case #13 in Table 1 ). E–H: Expression of tdTomato, indicating rAAV2/5, in NM cell bodies and fibers, also three weeks after injection (case #14 in Table 1 ). Two examples of labeled cell bodies are highlighted by white arrowheads. Note that in each row (A–C and E–G), the first three panels illustrate identical image frames but separately for 3 fluorescence channels; D and H show the corresponding merged images. Scale bars 50 ​μm. For further information about injection site, volume and expression time, see Table 1 .

Article Snippet: For fluorescent detection and imaging, a Nikon epifluorescence microscope (Nikon Eclipse Ni-E and the associated Nikon imaging software NIS Elements AR.4.30.02 64-Bit), as well as a fluorescence slide scanner (Axio Scan Z1, Zeiss, and the associated software ZEISS Efficient Navigation ZEN 2.6 blue edition) were used.

Techniques: Expressing, Injection, Labeling, Fluorescence

Journal: Current Research in Neurobiology

Article Title: Gene delivery to neurons in the auditory brainstem of barn owls using standard recombinant adeno-associated virus vectors

doi: 10.1016/j.crneur.2020.100001

Figure Lengend Snippet: Examples for the expression of serotypes rAAV2/1 and rAAV2/5 in nucleus laminaris . A–D: Expression of GFP, indicating rAAV2/1, in NM projection fibers ramifying within NL, three weeks after injection (case #13 in Table 1 ). E–H: Expression of tdTomato, indicating rAAV2/5, expressed in NM projection fibers ramifying and terminating within NL, also three weeks after injection (case #14 in Table 1 ). One example each of a labeled fiber (B) and terminals surrounding a (non-fluorescent) cell body (G) are highlighted by gray arrowheads, and also shown in higher-magnification insets. As in Fig. 2 , labeled fibers mostly appear as short fragments because their course was oblique to the sectioning plane. Note also that in each row (A–C and E–G), the first three panels illustrate identical image frames but separately for 3 fluorescence channels; D and H show the corresponding merged images. Scale bars 50 ​μm. For detailed information about injection site, volume and expression time, see Table 1 .

Article Snippet: For fluorescent detection and imaging, a Nikon epifluorescence microscope (Nikon Eclipse Ni-E and the associated Nikon imaging software NIS Elements AR.4.30.02 64-Bit), as well as a fluorescence slide scanner (Axio Scan Z1, Zeiss, and the associated software ZEISS Efficient Navigation ZEN 2.6 blue edition) were used.

Techniques: Expressing, Injection, Labeling, Fluorescence

Journal: Nutrients

Article Title: A Comparison of the Impact of Restrictive Diets on the Gastrointestinal Tract of Mice

doi: 10.3390/nu14153120

Figure Lengend Snippet: Restrictive diets have a small impact on goblet cells and mucus production in the GI tract, but affect intestine morphology. Histology sections of the colon ( A ) and jejunum ( B ) were stained with alcian blue periodic acid-Shiff (AB-PAS) to visualize goblet cells and mucus. The number of goblet cells per villi was counted in the jejunum sections ( C ). The mRNA expression of mucin (Muc2 and Muc13) genes was quantified in the mucosa of the jejunum ( D ) and proximal colon ( E ). Histological sections of jejunum were stained with hematoxylin-eosin ( F ), and the length of villi ( G ), depth of crypts ( H ), and thickness of muscularis externa ( I ) were quantified using ImageJ. Microscopy images were taken using Zeiss Axio Lab 5x HSF 22 microscope, Axiocam ERc 5s camera, and ZEN software (Zeiss efficient Navigation). The images were analyzed using ImageJ software. Statistical significance between experimental groups was evaluated using ANOVA and Tukey’s post hoc test. * p < 0.01; n = 6–8. Data are presented as the mean ± SEM. The scale bar in the last picture of A, B, and F is applied to all pictures.

Article Snippet: Images were taken using Axio Lab 5× HSF 22 microscope, Axiocam ERc 5s camera, and Zeiss Efficient Navigation (ZEN) software (Zeiss efficient Navigation) (Carl Zeiss Microscopy, Oberkochen, Germany).

Techniques: Staining, Expressing, Microscopy, Software

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