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: Neuron

Article Title: Neurotransmitter Switching Regulated by miRNAs Controls Changes in Social Preference

doi: 10.1016/j.neuron.2017.08.023

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-VGLUT1,2 , Synaptic Systems , Cat#135503; RRID: AB_2285905.

Techniques: Recombinant, Software

Journal: bioRxiv

Article Title: Imbalanced expression of clustered protocadherins in pre- and post-synaptic compartments of CA1 pyramidal cells during hippocampal development

doi: 10.1101/2023.04.15.536995

Figure Lengend Snippet: A : Images of Lucifer yellow-injected CA1 pyramidal cells. Scale bar: left, 200 μm; right, 5 μm. B : Image of pyramidal cell dendrites of P7 to 8-week old mice. Scale bar: 2 μm. C : Immunostaining for vGluT1 and magnified view (bottom) of the white square in the upper image. Scale bar: 100 μm (upper) and 20 μm (lower). D : Developmental stage-associated changes in spine density in the CA1 pyramidal cell radiatum region; mean ± SD. E : Developmental stage-associated changes in vGluT1 immunosignals in the CA1 stratum radiatum region; mean ± SD. F : Criteria for spinal morphology G : Developmental stage-associated changes in the proportion of each spine type.

Article Snippet: Similarly, slices were incubated overnight with rabbit anti-Lucifer yellow antibody and mouse anti-vesicular glutamate transporter-1 (vGluT1) antibody (0.4 μg/ml; sc-377425, Santa Cruz Biotechnology, Inc.) in the same primary solution.

Techniques: Injection, Immunostaining

Journal: bioRxiv

Article Title: Imbalanced expression of clustered protocadherins in pre- and post-synaptic compartments of CA1 pyramidal cells during hippocampal development

doi: 10.1101/2023.04.15.536995

Figure Lengend Snippet: A : cPcdhγCR immunosignals at P7. B : vGluT1 immunosignals. C : Merged image of cPcdhγ and vGluT1 expression. A’-C’ : Magnified view of the CA1 stratum radiatum regions. A”-C” : Magnified view of the upper images. D-F: Synaptic localization of cPcdhγCR immunosignals at P21. D : Green indicates dendrites and spines visualized using Lucifer yellow. cPcdhγ and vGluT1 were co-immunostained. The localization of cPcdhγ in synapses was categorized into four groups. E : Schematic representation of the immunosignal location categories. F : Proportion of cPcdhγ localization in the categories shown in E. Scale bar: 100 μm ( A-C ), 20 μm ( A’-C’ ), 1 μm ( A’’-C’’ ), 200 nm ( D ).

Article Snippet: Similarly, slices were incubated overnight with rabbit anti-Lucifer yellow antibody and mouse anti-vesicular glutamate transporter-1 (vGluT1) antibody (0.4 μg/ml; sc-377425, Santa Cruz Biotechnology, Inc.) in the same primary solution.

Techniques: Expressing

Journal: bioRxiv

Article Title: Imbalanced expression of clustered protocadherins in pre- and post-synaptic compartments of CA1 pyramidal cells during hippocampal development

doi: 10.1101/2023.04.15.536995

Figure Lengend Snippet: A-D’ : cPcdhγ (5 nm gold particles, red circles) and vGluT1 (15 nm gold particles) in the CA1 stratum radiatum. A, Pre-synaptic localization of cPcdhγ. cPcdhγ-immunoparticles were found near both the active zone ( A ) and the extra-synaptic area ( B ). C-C’ and D-D’ are images of paired replicas. C, D , Localization of cPcdhγ at postsynaptic sites. C’, D’, Localization of cPcdhγ at pre-synaptic sites. E, Proportion of cPcdhγ+ pre-synaptic sites identified using vGluT1 labelling. F, Number of cPcdhγ-immunoparticles in active zones and extra-active zones. G, Number of cPcdhγ-immunoparticles in pre- and postsynaptic faces of paired replicas. H, Proportion of cPcdhγ+ postsynapses in cPcdhγ+ pre-synapses. Scale bar: 200 nm.

Article Snippet: Similarly, slices were incubated overnight with rabbit anti-Lucifer yellow antibody and mouse anti-vesicular glutamate transporter-1 (vGluT1) antibody (0.4 μg/ml; sc-377425, Santa Cruz Biotechnology, Inc.) in the same primary solution.

Techniques:

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