Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: (A) Schematic diagram of the targeting strategy for the novel TMc- Nrg1 +/− mutant mice. Upper panel, wild type (WT) Nrg1 locus including 13 exons, shown as black boxes. The target vector, pKOS-53 with a LacZ/Neo cassette, was targeted to exon 9 of Nrg1 by homologous recombination. Lower panel, primer 0932-8, primer 0932-10, and primer Neo3a were used to distinguish WT alleles (346 bps) from mutant alleles (258 bps). (B) Mouse-tail DNA was submitted to PCR analysis. N: negative control. (C) The expression level of Nrg1 proteins in the brain of both WT and Nrg1 +/− mice. * P < 0.05 between two genotypes.

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Mutagenesis, Plasmid Preparation, Homologous Recombination, Negative Control, Expressing

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: The cognitive functions (mean + s.e.m.) of both male and female TMc- Nrg1 +/− mutant mice (black bars) and their wild-type littermate controls (WT, white bars) were evaluated in the Experiment 1 of this study. (A,B) In the object recognition task, a reduction in investigation time (s) of the novel object was observed in male, but not female, TMc- Nrg1 +/− mice. (C,D) In the contextual fear conditioning task, a reduction in freezing time percentage was observed in male, but not female, TMc- Nrg1 +/− mice. (E,F) In the cued fear conditioning task, a reduction in freezing time percentage (%) was found in male, but not female, TMc- Nrg1 +/− mice. In contrast, no significant genotype-dependent differences were found in the Morris water maze, prepulse inhibition task, or the social task in either males (G,I,K) or females (H,J,L) , respectively. Dash lines indicate chance level. * P < 0.05 between two genotypes.

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Mutagenesis, Inhibition

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: A summary of the basic behavioral phenotyping and statistical analyses (mean ± s.e.m) in the novel TMc- Nrg1 +/− ( Nrg1 +/− ) mutant mice and their wild-type (WT) littermates .

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Mutagenesis

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: Drug-induced behavioral alterations (mean + s.e.m.) in the novel TMc- Nrg1 +/− mice (black bars) and their wild-type controls (WT, white bars) ( n = 12 each group) of this study. (A,B) Total travel distances for the two groups of male and female mice during the 1-h saline baseline and the 1st and 2nd 1-h periods after the injection of MK-801 (0.25 mg/kg, i.p.). (C,D) Total travel distances (mean + s.e.m.) for the two groups of male and female mice during the 1-h saline baseline and during the 1st and 2nd 1-h periods after the injection of methamphetamine (MA, 2 mg/kg i.p.). (E) PTZ (60 mg/kg, s.c.)-induced seizure severity scores [from 0 (no response) to 4 (maximal seizure)] for both male and female mice. (F) Distributions of PTZ-induced seizure severity scores for male mice (%). * P < 0.05 between two genotypes.

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Saline, Injection

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: Neuromorphological analyses (mean ± s.e.m.) of GFP-labeled pyramidal neurons in the hippocampi of male and female TMc- Nrg1 +/− ( Nrg1 +/− ) mice and their wild-type (WT) littermate controls .

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques:

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: The neurochemical analysis of the expression of GABAergic markers in the hippocampi of the novel TMc- Nrg1 +/− mutant mice (black bars) and their wild-type controls (WT, white bars) determined by Western blot. (A,B) Representative Western blots of hippocampal tissue (30 μg/lane) with GAD67 (67 kDa), calretinin (29 kDa), parvalbumin (12 kDa), and GAPDH (37 kDa) from males and females. (C–H) Quantification of the data revealed reductions in GAD67 and parvalbumin expression levels in the hippocampi of male (C,E,G) , but not female (D,F,H) , TMc- Nrg1 +/− mice. Data are presented as the mean + s.e.m. * P < 0.05 between two genotypes.

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Expressing, Mutagenesis, Western Blot

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: The effect of chronic valproate treatment on the rescue of cognitive deficits in male wild-type (WT, white bars) and TMc- Nrg1 +/− mutant (black bars) mice . Three cognitive tasks, including (A) an object recognition task, (B) a contextual fear conditioning task, and (C) a cued fear conditioning task, were conducted. Either valproate (1.5 mmol/kg, s.c.) or 0.9% saline was injected twice a day for 17 days. The object recognition task was conducted on day 16,and the contextual/cued tests of fear conditioning were performed on day 17. (D) The expression of GAD67 in the hippocampus of both WT and TMc- Nrg1 +/− mice after chronic injections of either saline or valproate. Data are presented as the mean + s.e.m. * P < 0.05 between two genotypes.

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Mutagenesis, Saline, Injection, Expressing

Journal: Frontiers in Behavioral Neuroscience

Article Title: Distinct phenotypes of new transmembrane-domain neuregulin 1 mutant mice and the rescue effects of valproate on the observed schizophrenia-related cognitive deficits

doi: 10.3389/fnbeh.2014.00126

Figure Lengend Snippet: Summary of the behavioral phenotypes and comparison of the different lines of Nrg1 -related mutant mice .

Article Snippet: The expression of Nrg1 protein in the cerebral cortex of the novel TMc- Nrg1 +/− mutant mice was further validated using Western blot with a Nrg1 C-terminal antibody (Neuregulin-1α/β1/2, 1:500, sc348, Santa Cruz Biotechnology).

Techniques: Comparison, Mutagenesis, Activity Assay, Inhibition, Transgenic Assay

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) CD11c + cells are increased in the lungs of atopic mice. Flow cytometry of lung cell suspension showing frequency (left) and cell number (right) at day 3 post inoculation (PI) high dose SeV ( B ) Adoptive transfer of CD11c + cells isolated from NA or atopic mice into naïve mice 24 h before inoculation with high dose SeV delays but does not prevent mortality; n=4 per group. ( C ) Transcriptomic (RNAseq) comparison between FACS isolated lung CD11c + cells from atopic and NA mice identifies several disparately expressed gene products, including Nrg1 (n=4 per group). Selected gene products shown. ( D ) NRG1 is markedly increased in atopic mouse lung (“tissue”), BAL, and supernatant from 1 x10 6 atopic lung CD11c + cells (“CD11c sup”) cultured for 24 h. ( E ) The scRNA (10x Platform) tSNE coordinates show CD14 + monocytes and dendritic cell (“DC”) sub-populations expressing NRG1 in peripheral blood cells of healthy human donors.*p<0.05, **p<0.01, ****p<0.0001

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: Flow Cytometry, Suspension, Adoptive Transfer Assay, Isolation, Comparison, Cell Culture, Expressing

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) NRG1(1ng to 1000ng) i.n. (in 30µL) given daily to naïve mice for 5d before inoculation with high dose SeV reduces viral mortality; n=4 per group (1ng, 10ng 1000ng & PBS), n=8 per group (100 ng and 500 ng). ( B ) Ratio of EBD in the BAL to that in the lung shows reduced EBD in NRG1 treated mice on day 8 PI SeV. n≥8 per group, median ± IQR shown, Mann-Whitney U test.

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) Adding NRG1 to hBEC inoculated with rgRSV (left) and mTEC inoculated with GFP-SeV (right) reduces spread of infection. Representative images shown. ( B ) Quantification of (A) for rgRSV and hBEC and ( C ) for GFP-SeV and mTEC. GFP positive cells quantified by ImageJ. Representative images from ≥3 separate experiments. *p<0.05, **p<0.01. ( D ) Transcriptomic analysis of hBEC cultures treated with NRG1 (100 ng) on the basolateral side of the Transwell for 5 days and inoculated with RSV (4000 pfu). RNA was isolated 48 h PI RSV and qRT-PCR performed using a custom Prime PCR array plate: (i) Transcripts in which NRG1 treatment reduced gene expression from that seen in RSV infected cells. (ii) Transcripts with low level expression that show small but significant change in expression relative to naïve control with NRG1 alone or genes significantly increased with RSV but whose expression levels were not affected by NRG1. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, n=3.

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: Infection, Isolation, Quantitative RT-PCR, Expressing, Control

Journal: bioRxiv

Article Title: Regionalized cell and gene signatures govern oesophageal epithelial homeostasis

doi: 10.1101/2024.02.21.581361

Figure Lengend Snippet: (A-B) Circular plot displaying number of interactions between proximal fibroblasts and proximal epithelial cells (A) and between distal fibroblasts and distal epithelial cells (B). Epithelial cells were aggregated into basal, suprabasal, and proliferative cells based on the annotation of subclustered epithelial cells. (C) Circular plot of differential interaction strengths in cell-cell communication between fibroblasts and epithelial cells comparing datasets containing proximal epithelial cells and fibroblasts and distal epithelial cells and fibroblasts, respectively. (D) Information flow (sum of communication probabilities among all pairs of cell groups) of all significantly altered signalling pathways comparing proximal and distal derived cells. (E, F) Heatmaps of selected incoming (blue) (E) and outgoing (green) (F) signalling pathways of regional oesophageal fibroblasts and epithelial cells. (G) Violin plots displaying regional gene expression of expressed WNT ligands in epithelial basal cells. (H) In situ hybridization of Axin2 (red) and Wnt5a (green) on cross-sections of the proximal and distal oesophagus. Sections are counterstained with E-CADHERIN (ECAD, white) and DAPI (blue). Scale bar = 20 µm. Dotted line marks the epithelial-stromal border. (I-J) Violin plots displaying regional gene expression of selected BMP ligand (I) and Igf1 (J) in fibroblasts. (K-L) Violin plot displaying regional gene expression of Igf1r (K) and Nrg1 (L) in basal and suprabasal epithelial cells.

Article Snippet: All supplemented media are based on ENR lo medium and contain either, 250ng/ml recombinant IGF1 (Peprotech, #100-11), 250ng/ml recombinant WNT5A (R&D systems, #645-WN-010), 100ng/ml BMP4 (R&D systems, #5020-BP-010), 5ng/ml IL-17A (R&D systems, #421-ML-025/CF), or 100ng/ml recombinant NRG1 (Biotechne, #9875-NR-050).

Techniques: Derivative Assay, Expressing, In Situ Hybridization

Journal: bioRxiv

Article Title: Regionalized cell and gene signatures govern oesophageal epithelial homeostasis

doi: 10.1101/2024.02.21.581361

Figure Lengend Snippet: (A) Information flow (sum of communication probabilities among pairs of cell groups) of selected signalling pathways comparing datasets encompassing all proximal and distal cells within the single-cell dataset. (B, D, F, H) Comparison of the outgoing and incoming interactions strengths between proximal-distal fibroblasts and proximal-distal epithelial cells, focusing on (B) WNT-, (D) BMP-, (F) IGF- and (H) NRG-signalling. (C) In situ hybridization for Wnt4 (red), enriched in the epithelium. (E) In situ hybridization for Bmp7 (red). White arrows mark Bmp7 -positive stromal fibroblasts in close contact with the epithelium. (G) In situ hybridization for Igf1 (red), enriched in the distal, compared to the proximal, stroma. (I) In situ hybridization for Nrg1 (red), expressed in the proximal, but not distal, epithelium. All in situ hybridizations (C, E, G, and I) are performed on cross-sections of the proximal and distal oesophagus and counterstained with E-CADHERIN (ECAD, white) and DAPI (blue). Dotted lines mark the epithelial-stromal border. Scale bar = 20µm.

Article Snippet: All supplemented media are based on ENR lo medium and contain either, 250ng/ml recombinant IGF1 (Peprotech, #100-11), 250ng/ml recombinant WNT5A (R&D systems, #645-WN-010), 100ng/ml BMP4 (R&D systems, #5020-BP-010), 5ng/ml IL-17A (R&D systems, #421-ML-025/CF), or 100ng/ml recombinant NRG1 (Biotechne, #9875-NR-050).

Techniques: Comparison, In Situ Hybridization, In Situ

Journal: bioRxiv

Article Title: Regionalized cell and gene signatures govern oesophageal epithelial homeostasis

doi: 10.1101/2024.02.21.581361

Figure Lengend Snippet: (A) Quantification of proximal and distal organoid forming efficiency (OFE) comparing control medium and medium containing 100ng/mL BMP4. (B) Quantification of organoid size in control medium and medium supplemented with 250ng/mL IGF1. (C-D) Quantification of organoid forming efficiency (C) and size (D) in control medium and medium supplemented with 100ng/mL recombinant NRG1. (E) Quantification of organoid forming efficiency in the presence of 250ng/mL WNT5A. All quantifications were performed at day 6 of culture. For size quantifications each dot presents average size of all organoids. n=3-5. (F) Graphical summary illustrating the regionalized oesophageal stromal and epithelial cell distributions as well as signalling gradients. (A-E) Two-sided ratio paired t-test. ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: All supplemented media are based on ENR lo medium and contain either, 250ng/ml recombinant IGF1 (Peprotech, #100-11), 250ng/ml recombinant WNT5A (R&D systems, #645-WN-010), 100ng/ml BMP4 (R&D systems, #5020-BP-010), 5ng/ml IL-17A (R&D systems, #421-ML-025/CF), or 100ng/ml recombinant NRG1 (Biotechne, #9875-NR-050).

Techniques: Recombinant