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: Frontiers in Neuroanatomy

Article Title: A Computational Synaptic Antibody Characterization Tool for Array Tomography

doi: 10.3389/fnana.2018.00051

Figure Lengend Snippet: Schematic diagram of input datasets. (A) The target protein, gephyrin, is a postsynaptic protein at inhibitory synapses, expected to be adjacent to GAD, a presynaptic protein abundant at inhibitory synapses. The set of squares represents a stack of images from serial ultrathin sections from mouse neocortex, double labeled with a gephyrin candidate antibody (brown dots) and a previously validated antibody against GAD (red dots). The large blue blobs represent DAPI, a marker for cell nuclei. (B) Identical setup, but with Bassoon (a presynaptic protein present in excitatory synapses) as the target protein. In this example, the tissue is labeled with two reference antibodies to excitatory synapses, the presynaptic protein VGluT1 (purple dots) and the postsynaptic protein PSD-95 (red dots). (C) Different combinations of puncta are detected on sections ( z = 0 to z = 3) through a synapse. The candidate and the reference antibody can be present alongside each other on the same section (a), they can lie adjacent in the z-direction (b), or they can be adjacent both in the same section and across multiple sections (c). (D) Identical setup as C with two reference antibodies depicted.

Article Snippet: VGluT1 , Mouse , NeuroMab 75-066 , RRID:AB_2187693.

Techniques: Labeling, Marker

Journal: Frontiers in Neuroanatomy

Article Title: A Computational Synaptic Antibody Characterization Tool for Array Tomography

doi: 10.3389/fnana.2018.00051

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: VGluT1 , Mouse , NeuroMab 75-066 , RRID:AB_2187693.

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: A Computational Synaptic Antibody Characterization Tool for Array Tomography

doi: 10.3389/fnana.2018.00051

Figure Lengend Snippet: Results from pairwise antibody comparisons.

Article Snippet: VGluT1 , Mouse , NeuroMab 75-066 , RRID:AB_2187693.

Techniques: Standard Deviation

Journal: Frontiers in Neuroanatomy

Article Title: A Computational Synaptic Antibody Characterization Tool for Array Tomography

doi: 10.3389/fnana.2018.00051

Figure Lengend Snippet: Five antibodies evaluated at different concentrations.

Article Snippet: VGluT1 , Mouse , NeuroMab 75-066 , RRID:AB_2187693.

Techniques: Standard Deviation

Journal: Frontiers in Neuroanatomy

Article Title: A Computational Synaptic Antibody Characterization Tool for Array Tomography

doi: 10.3389/fnana.2018.00051

Figure Lengend Snippet: Summary of candidate antibody comparisons.

Article Snippet: VGluT1 , Mouse , NeuroMab 75-066 , RRID:AB_2187693.

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: A Computational Synaptic Antibody Characterization Tool for Array Tomography

doi: 10.3389/fnana.2018.00051

Figure Lengend Snippet: Comparison of multiple candidate antibodies using two reference synaptic antibodies. Left: Bassoon with PSD-95 and VGluT1. Right: IRSp53 with PSD-95 and VGluT1. Expert ranking is color coded: green—best, orange—unclear, red—fail. Compare with Figure .

Article Snippet: VGluT1 , Mouse , NeuroMab 75-066 , RRID:AB_2187693.

Techniques: Comparison

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: 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 neurology

Article Title: Dual Motor Cortex and Spinal Cord Neuromodulation Improves Rehabilitation Efficacy and Restores Skilled Locomotor Function in a Rat Cervical Contusion Injury Model

doi: 10.1016/j.expneurol.2021.113715

Figure Lengend Snippet: Proprioceptive afferent terminations on CTB-labelled ECR motoneurons. A, B) Confocal micrographs of VGlut1 on motoneurons. A: Rehab-only, B: Stim+rehab. Rehab-only group has more VGlut-1 terminations contacting the boarder and proximal dendrites of ECR motoneurons. VGlut1 on motoneurons is a marker for PA synaptic terminals. Figures illustrate greater monosynaptic PA-motoneuron connections after rehab-alone, whereas stim+rehab seems to attenuate such synaptic formation. Scale bars: 50 μm. C) Bar graph showing rehab-only group has significantly more VGlut-1 per motoneurons (boarder and proximal dendrites) compared to stim+rehab.

Article Snippet: Vesicular glutamate transporter 1 (VGlut1) was localized using a primary anti-VGlut1 antibody (1:10; NeuroMab) and a secondary donkey anti-mouse conjugated to Cy5.

Techniques: Marker

Journal: Journal of molecular neuroscience : MN

Article Title: Interactions of Antibodies to the Gram-Negative Gastric Bacterium Helicobacter pylori with the Synaptic Calcium Sensor Synaptotagmin 5, Correlate to Impaired Vesicle Recycling in SiMa Human Neuroblastoma Cells.

doi: 10.1007/s12031-020-01670-0

Figure Lengend Snippet: Fig. 3 Western blot analysis of the cross-reactivity of antibodies directed to Helicobacter pylori (α-HPy) and Campylobacter jejuni (α-CJe) with different protein samples as provided by commercial HEK-293 overexpression lysates. a Cross-reactivity of α-HPy as revealed by a distinct immunopositive band can be observed for Syt5 and Vglut1 (Slc17a7), whereas Stmn4 and Ncan reveal no such band. Also a control lysate of non-transfected HEK293 cells, as well as an overexpression lysate of Srf is negative. b For α-CJe cross-reactivity as revealed by a distinct immunopositive band can be observed for Syt5 and Vglut1, whereas also in this case Stmn4 and Ncan reveal no such band. Again a control lysate of non-transfected HEK293 cells, as well as an overexpression lysate of Srf is negative. A corresponding negative control incubated with secondary antibodies only is shown in Supplementary Fig. 2

Article Snippet: H um a n SLC17A7-transfected HEK293 overexpression lysate (Origene, cat. no. LY412521).

Techniques: Western Blot, Over Expression, Control, Transfection, Negative Control, Incubation