Journal: bioRxiv

Article Title: Microglia undergo transcriptional, translational and functional adaptations to dark and light phases in laboratory mice

doi: 10.1101/2023.11.17.567571

Figure Lengend Snippet: Microglia display increased synaptic pruning during the active phase In the adult hippocampus. A Schematic representation of the experimental setup. Whole hippocampi were retrieved from perfused adult male mice either 4h post lights-off (active phase) or 4h post lights-on (sleep phase) for CD11b cell enrichment and subsequent flow cytometry analysis of vGlut1-posive cells. B Heatmap showing the differential expression (in average transcripts per million, TPM), for selected genes associated with microglial phagocytic functions from the total-RNA-seq of microglia 4h post lights-off or 4h post lights-on ( , 1 a.m. versus 1 p.m., Only genes that were significantly differentially expressed are represented, adj. p -value < 0.05) C Scatter plot showing the difference in percentage of vGlut1-positive microglia cells between the active and sleep phase, 4h post lights-off and on respectively, (unpaired t-test, t(5.84) = 4.84, p = 0.0031, N = 6/group). D Histogram showing the microglial vGlut1-PE mean fluorescence intensity normalized to spleen cells as negative control (lower panel). E Scatter plot showing the area under the curve (Au) for the microglial vGlut1 mean fluorescence intensity during the active (4h after light-off) and sleep (4h post light-on) phases, normalized to spleen cells as negative controls (unpaired t-test, t(10) = 2.98, p = 0.014, N = 6/group). F Heatmap showing differential expression (in average transcripts per million, TPM), for selected genes coding for chemokines (from the total-RNA-seq of microglia 1 p.m. versus 1 a.m.). Only genes that were significantly differentially expressed are represented (adj. p -value < 0.05). G Scatter plot comparing the percentage of CD45 hi cell population in the hippocampi of adult mice 4h after light-off (N = 17) and 4h post light-on (N = 13, unpaired t-test, t(28) = 3.08, p = 0.0046). Error bars represent the mean ± standard deviation. * p < 0.05, ** p < 0.01. Images created with Biorender.com.

Article Snippet: Following fixation, intracellular staining for vGLUT1 (1/200, Miltenyi Biotec, #130-120-764, 1h in 1x BD Permeabilization Buffer) was immediately performed.

Techniques: Flow Cytometry, Quantitative Proteomics, RNA Sequencing, Fluorescence, Negative Control, Standard Deviation

Journal: bioRxiv

Article Title: Microglia undergo transcriptional, translational and functional adaptations to dark and light phases in laboratory mice

doi: 10.1101/2023.11.17.567571

Figure Lengend Snippet: Gating strategy for the flow cytometry analysis of vGLUT1- positive inclusions within hippocampal microglia cells sorted during wither the active or the sleep phase. Supplementary Figure S6: A -log(FDR) values for biological processes associated with the genes deregulated in response to LPS in function of time of injection. B Unbiased Tmod enrichment analysis for reactome gene sets for the LPS response genes regulated by time of injection.

Article Snippet: Following fixation, intracellular staining for vGLUT1 (1/200, Miltenyi Biotec, #130-120-764, 1h in 1x BD Permeabilization Buffer) was immediately performed.

Techniques: Flow Cytometry, Injection

Journal: Molecular Brain

Article Title: Exploration of schizophrenia-related behavioral and molecular abnormalities in a mutant mouse model with a mutation in the TVV motif of the ErbB4 gene

doi: 10.1186/s13041-025-01238-2

Figure Lengend Snippet: ErbB4 mutation disrupts key proteins of postsynaptic signaling. ErbB4 is increased in homo mice while phosphorylated ErbB4 is decreased and less activated compared to abundance of total protein amount ( A , B , Student’s t-test, p < 0.0001, p < 0.001, and p < 0.0001 respectively). In contrast, the total amount of PSD-95 is decreased (Student’s t-test, p < 0.05), but the remaining pool is highly activated in mutant mice ( A , B , Student’s t-test, p < 0.05). Mutation also caused hypofunction of NMDAR2A subunit ( A , C ). NMDAR2A is highly decreased (Student’s t-test, p < 0.001) but hyperactivated in mutant homo mice (Student’s t-test, p < 0.01). In contrast, NMDAR1 subunit was not affected which highlights the mutation caused NMDAR subunit specific defects. The GABAergic system is also selectively disrupted ( A , D ). GAD65 a global brain GABAergic maker and VGLUT1 were not affected (Student’s t-test, p > 0.05) but the GAD67 isoform is specifically highly decreased (Student’s t-test, p < 0.01) in mutant homo mice. 60 µg protein, N = 3 and n = 6 in all experiments, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001, N = number of mice, and n = number of samples

Article Snippet: The membranes were blocked with quick blocking buffer (P0252 Beyotime) for 15 min and incubated at 4 °C overnight with the following primary antibodies: rabbit anti-ErbB4 (1:1000, Cat No. 4795, CST), mouse anti-PSD-95 (1:1000, Cat No. MAB1598, Sigma Aldrich), mouse anti-GAPDH (1:5000, Cat No. HC301, TransGen), rabbit anti-phospho-phospho-PSD-95 (1:1000, Cat. No. 45737, CST), rabbit anti-phospho-ErbB4 (1:1000, Cat No. 4757, CST), rabbit anti-NMDA receptor 2 A (1:1000, Cat. No. 4205, CST), rabbit anti-phospho-NMDA receptor 2 A (1:1000, Cat. No. 4206, CST), rabbit anti-GAD67 (1: 2000, Cat. No. ab239372, Abcam), rabbit anti-GAD67 (1: 1000, Cat. No. ab97739, Abcam), mouse anti-NMDAζ1 (1:300, Cat. No. sc-518053, Santa Cruz Biotechnology), and mouse VGLUT1 (1:500, Cat. No. NBP2-59329, Novus Bio).

Techniques: Mutagenesis

Journal: Neuropsychopharmacology Reports

Article Title: Preliminary analysis of hippocampus synaptic apoptosis and microglial phagocytosis induced by severe restraint stress

doi: 10.1002/npr2.12298

Figure Lengend Snippet: We acquired images from CA3 stratum lucidum region in control and stressed male C‐57 BL6 mice using confocal laser scanning microscopy, subsequently for analysis converted and binarize them using image converter/analyzer. Scale bar, 5 μm. (A) representative converted (left side) and binarized (right side) images in control and stressed mice. (B) ratios of the area where VGLUT1 co‐localizes with caspase 3 to the VGLUT1+ area.10‐d water immersion restraint stress significantly increase the value (control: n = 4 animals, stress: n = 3 animals, t5 = 3.0265, P = .0292). Error bars, SD

Article Snippet: For evaluating local synaptic apoptosis, sections were double stained with rabbit anti‐cleaved caspse‐3 (1:300; Cell Signaling, #9664) and guinea pig anti‐ vesicular glutamate transporter1 (VGLUT1) antibody (1:2000; synaptic systems, #135304) for 6 days at 4°C.

Techniques: Control, Confocal Laser Scanning Microscopy

Journal: Neuropsychopharmacology Reports

Article Title: Preliminary analysis of hippocampus synaptic apoptosis and microglial phagocytosis induced by severe restraint stress

doi: 10.1002/npr2.12298

Figure Lengend Snippet: (A) Representative 3D reconstruction images obtained from CA3 stratum lucidum region in control and stressed male C‐57 BL6 mice. Iba1+ microglia and LAMP1+ structures are visualized at left side. Iba1+ microglia, LAMP1+ structures and VGLUT1+ structures are visualized at right side. Scale bar, 5 μm. (B) Ratios of the volume of the LAMP1+ structures contained in the Iba1+ microglia to the volume of the Iba1+ microglia. There was no significant difference between control and stressed groups in the ratio (control: n = 4 animals, stress: n = 3 animals, t5 = 0.8946, P = .412). Error bars, SD . (C) Ratios of the volume of engulfed VGLUT1 within LAMP1+ structures contained in the Iba1+ microglia to the volume of the Iba1+ microglia. There was no significant difference between control and stressed groups in the ratio (control: n = 4 animals, stress: n = 3 animals, t5 = 0.4414, P = .6773). Error bars, SD

Article Snippet: For evaluating local synaptic apoptosis, sections were double stained with rabbit anti‐cleaved caspse‐3 (1:300; Cell Signaling, #9664) and guinea pig anti‐ vesicular glutamate transporter1 (VGLUT1) antibody (1:2000; synaptic systems, #135304) for 6 days at 4°C.

Techniques: Control

Journal: Ecotoxicology and environmental safety

Article Title: Influence of bifenthrin exposure at different gestational stages on the neural development.

doi: 10.1016/j.ecoenv.2023.115365

Figure Lengend Snippet: Fig. 5. Effect of BF treatment at GD 16-B on the protein levels of VGluT1, VGAT, and NMDA receptors in mouse hippocampi at 6-week-old. (A) Repre sentative western blot images of VGluT1, VGAT, NR1, NR2A and NR2B expression in the mouse hippocampi at 6-week-old. These western blot images are from one membrane. Taking β-Tubulin as loading control. (B) & (C) Quantization of the protein levels of VGluT1, VGAT, VGluT1/VGAT, NR1, NR2A and NR2B of the male mouse (B) and female mouse hippocampi (C) at 6-week-old. N = 4 (litter). The bands in each column of Fig. 5A are from the same pup. Unpaired Student’s t-test was used to analyze the statistical significance. * * P < 0.01 vs. respective sex control.

Article Snippet: Primary antibodies against VGluT1 (#CL2754), VGAT (#131003), NMDA Receptor 2B (NR2B, GluN2B, #21920–1-AP) were bought from Novus Biologicals (Littleton, CO, USA), synaptic systems (Gottingen, Germany), and Proteintech (Chicago, IL, USA) respectively.

Techniques: Western Blot, Expressing, Membrane, Control

Journal: Experimental Neurobiology

Article Title: Sensory Stimulation-dependent Npas4 Expression in the Olfactory Bulb during Early Postnatal Development

doi: 10.5607/en23037

Figure Lengend Snippet: Information on primary antibodies used in western blot

Article Snippet: Anti- VGLUT1 , Atlas antibodies , Amab91041 , WB 1:1000.

Techniques: Western Blot, Concentration Assay

Journal: Experimental Neurobiology

Article Title: Sensory Stimulation-dependent Npas4 Expression in the Olfactory Bulb during Early Postnatal Development

doi: 10.5607/en23037

Figure Lengend Snippet: Effect of unilateral naris occlusion on OB interneurons during postnatal olfactory bulb development. (A) Immunoblot of GAD67 and VGLUT1 in P14 olfactory bulbs isolated from the open and closed sides of the naris. (B) The relative levels of GAD67 and VGLUT1 normalized by β-actin level (n=11, ***p<0.001). (C) Immunoblots of tyrosine hydroxylase (TH) and calretinin (CALR) normalized by β-actin level in P14 olfactory bulbs isolated from the open and closed sides of the naris. (D) The relative levels of TH and CALR were normalized by β-actin level (n=11, **p<0.01, ****p<0.0001).

Article Snippet: Anti- VGLUT1 , Atlas antibodies , Amab91041 , WB 1:1000.

Techniques: Western Blot, Isolation