c fos nuclei  (Oxford Instruments)

Journal: Neurobiology of Stress

Article Title: Post-weaning social isolation in male mice leads to abnormal aggression and disrupted network organization in the prefrontal cortex: Contribution of parvalbumin interneurons with or without perineuronal nets

doi: 10.1016/j.ynstr.2023.100546

Figure Lengend Snippet: PWSI-induced abnormal aggression is associated with subregion-specific neuronal hyperactivation in the mPFC and disrupted co-activation patterns across mPFC and OFC subregions. A , Experimental design. Upon weaning, mice were housed individually (PWSI) or were housed in groups of four (social). After reaching adulthood mice were subjected to a behavioral test battery. On PN109, mice were perfused under baseline conditions (BASE) or 90 min after the aggressive interaction (AGGR) followed by c-Fos immunostaining to investigate the impact of PWSI on the neuronal activation of the PFC. B–C, Aggressive interaction induced an mPFC-specific enhanced c-Fos expression in PWSI mice compared to social mice. B , Schematic drawing showing the Bregma level of the investigated orbitofrontal (OFC) subregions and bar graph showing the number of c-Fos immuno-positive nuclei within the OFC, including the ventral orbitofrontal (VO) and medial orbitofrontal cortices (MO). C , Schematic drawing showing the Bregma level of the investigated medial prefrontal (mPFC) subregions and bar graph showing the number of c-Fos immuno-positive nuclei within the mPFC, including the anterior cingulate (ACC), the prelimbic (PrL) and the infralimbic cortices (IL). D-G , PWSI induced disrupted functional connectivity across PFC subregions. Cross-correlation and network connectivity analysis of c-Fos expression in experimental groups, computed as covariance across subjects to reveal interactions between PFC subregions. The top row represents the complete set of interregional correlations in each experimental group, whereas the bottom row depicts node graphs where the nodes represent brain regions and the width of connections between nodes indicate the higher correlation coefficients. Connectivity matrices and node graphs representing social BASE ( D ), PWSI BASE ( E ), social AGGR ( F ) and PWSI AGGR ( G ) groups. Colors indicate Pearson correlation coefficients (scale, right) and labels within squares correspond to p values of correlations. All data are represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. ACC, anterior cingulate cortex; AGGR, aggressive interaction; BASE, baseline condition; IL, infralimbic cortex; MO, medial orbitofrontal cortex, mPFC, medial prefrontal cortex; OFC, orbitofrontal cortex; PN, postnatal day; PrL, prelimbic cortex, RI1, first resident-intruder test; RI2, second resident-intruder test; VO, ventral orbitofrontal cortex. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Microscopic images were digitized by an Olympus CCD camera using a 20x magnification lens and c-Fos immunoreactive nuclei were counted using Fiji as described previously ( ).

Techniques: Activation Assay, Immunostaining, Expressing, Functional Assay

Journal: Neurobiology of Stress

Article Title: Post-weaning social isolation in male mice leads to abnormal aggression and disrupted network organization in the prefrontal cortex: Contribution of parvalbumin interneurons with or without perineuronal nets

doi: 10.1016/j.ynstr.2023.100546

Figure Lengend Snippet: Overview of primary antibodies or reagents used in this study.

Article Snippet: Microscopic images were digitized by an Olympus CCD camera using a 20x magnification lens and c-Fos immunoreactive nuclei were counted using Fiji as described previously ( ).

Techniques:

Journal: Neurobiology of Stress

Article Title: Post-weaning social isolation in male mice leads to abnormal aggression and disrupted network organization in the prefrontal cortex: Contribution of parvalbumin interneurons with or without perineuronal nets

doi: 10.1016/j.ynstr.2023.100546

Figure Lengend Snippet: PWSI-induced abnormal aggression correlates with mPFC-specific heightened activity of PV+PNN+ neurons. For the experimental timeline see A. A-D , Aggressive interaction increased the activity of PV + neurons in the OFC. A , Representative single plane confocal image showing activated (c-Fos-immunpositive, yellow) parvalbumin neurons (parvalbumin-immunpositive, magenta) surrounded by PNNs (stained with WFA, cyan) from the OFC of a PWSI mouse. B , Aggressive interaction (AGGR) increased the number of activated PV+ neurons compared to baseline, i.e. resting controls (BASE). Graphs show the density of activated parvalbumin neurons (PV+cFos+ double immunopositive) in the OFC. C-D, Aggressive interaction increased the activation of both PV+PNN- ( C) and PV+PNN+ ( D) neuronal populations in the OFC of social and PWSI mice. E-H , PWSI induced hyperactivation of PV + neurons in the mPFC following aggressive interaction. E , Representative single plane confocal image showing activated (c-Fos+, yellow) parvalbumin neurons (PV+, magenta) with (stained with WFA, cyan) and without PNNs from the mPFC of a PWSI mouse. F , Aggressive interaction-specific significant increase in the density of activated PV+ neurons (PV+cFos+, double immunopositive) in the mPFC of PWSI mice compared to social mice. G-H, The density of activated PV+PNN- neurons ( G) and PV+PNN+ neurons ( H) in the mPFC revealed a PWSI-specific hyperactivation of PV+PNN+ neurons. I-J , PWSI altered correlations between behavioral variables during aggressive interaction and activation patterns of distinct PV + neuronal populations in the PFC. I, Spearman correlation matrices for social mice (upper panel) and PWSI mice (lower panel) showing relationships between OFC PV+ neuron activation and behavior during AGGR. J , Spearman correlation matrices for social mice (upper panel) and PWSI mice (lower panel) showing relationships between mPFC PV + neuron activation and behavior during AGGR. Each square shows the Spearman's correlation coefficient as calculated by correlating mean densities of activated PV+ neurons, PV+PNN- neurons or PV+PNN+ neurons with hallmark aggression features shown in . Colors indicate Spearman correlation coefficients, and labels within squares correspond to p values of correlations. All data are represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005, AGGR, aggressive interaction; BASE, baseline i.e. resting conditions; mPFC, medial prefrontal cortex; OFC, orbitofrontal cortex; PV, parvalbumin; PV+PNN-, parvalbumin neurons without perineuronal nets; PV+PNN+, parvalbumin neurons with perineuronal nets. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Microscopic images were digitized by an Olympus CCD camera using a 20x magnification lens and c-Fos immunoreactive nuclei were counted using Fiji as described previously ( ).

Techniques: Activity Assay, Staining, Activation Assay

Journal: PLoS ONE

Article Title: B 0 AT2 (SLC6A15) Is Localized to Neurons and Astrocytes, and Is Involved in Mediating the Effect of Leucine in the Brain

doi: 10.1371/journal.pone.0058651

Figure Lengend Snippet: (A) Cumulative food intake of regular chow showed no difference during the 6 days of measurements. Data on the graphs are shown as food intake/body weight (BW) (g) ± SD, for Slc6a15 KO mice (−/−) and WT mice (+/+) (n = 19–20/group). (B and C) Food intake was analyzed by dividing the actual food intake for each animal with the weight of the animal 0–2, 0–4 and 0–24 hours after leucine (Leu) or valine (Val) injection in Slc6a15 KO and WT mice (n = 20/group). WT mice given leucine showed a significant (p = 0.0268) reduction of food intake 2 hours after injection. Valine given WT mice showed a decreased but not significant food intake, when measured 2 hours after injection. The food intake after 4 hours or 24 hours was not significantly different between the two groups. (D and E) Number of activated neurons ± SEM (c-Fos positive cells) in food related brain regions after leucine or valine injections in Slc6a15 KO and WT mice (n = 6/group). WT mice had a significant leucine and valine induced increase (p Leu = 0.0350, p Val = 0.0319) of c-Fos expression in VMH. Both leucine and valine injection gave trends of reduced activation of c-Fos in Arc in WT mice. WT mice had a non-significant reduction of c-Fos expression in dorsal medial hypothalamus (DMH) after leucine injection, a reduction not seen after valine injection. No difference in c-Fos activation was seen in paraventricular nucleus of hypothalamus (PVN) after leucine or valine injection. (F and G) Number of activated pS6 positive neurons ± SEM (c-Fos and pS6 positive cells) in brain regions after leucine or valine injections in Slc6a15 KO and WT mice (n = 5–6/group). Valine induced significant reduction (p = 0.0096) of activated neurons in the mTOR pathway in VMH in WT mice. Leucine injection non-significantly increased the number of c-Fos and pS6 positive cells in VMH in WT mice, reduced the number of cells co-labelled with c-Fos and pS6 in arcuate nucleus of hypothalamus (Arc) in WT mice, while valine gave no difference between the two groups. Both leucine and valine injection gave trends of increased activation in PVN in WT mice. Leucine injections decreased while valine injections increased the trends of activation of c-Fos and pS6 positive cells in DMH in WT mice. Asterisks denote significance of Student's t-test for difference of means (*p<0.05, **p<0.01, *** p<0.001).

Article Snippet: Brain regions of interest were annotated and c-Fos positive nuclei (leucine experiment II) or co-localization of c-Fos and pS6 positive cells (leucine experiment III) were manually quantified, using the Pannoramic viewer software v.1.14 (3DHistech, Hungary), based on 5–6 mice/group.

Techniques: Injection, Expressing, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: Constitutively active microglial populations limit anorexia induced by the food contaminant deoxynivalenol

doi: 10.1186/s12974-022-02631-7

Figure Lengend Snippet: Immunohistochemistry conditions

Article Snippet: Counting of c-Fos positive nuclei was performed on photomicrographs acquired using a tenfold lens with a DXM 1200 Camera (Nikon) coupled to ACT-1 software.

Techniques: Immunohistochemistry, Blocking Assay, Plasmid Preparation

Journal: Journal of Neuroinflammation

Article Title: Constitutively active microglial populations limit anorexia induced by the food contaminant deoxynivalenol

doi: 10.1186/s12974-022-02631-7

Figure Lengend Snippet: PLX3397-induced microglia deletion exacerbates DON-induced anorexia. A Representative images of IBA1 staining in the dorsal vagal complex (DVC) of animal fed for 3 weeks with normal chow (NC) or food supplemented with PLX3397 (290 mg/kg chow). Scale bar: 100 µm. B–C Cumulative food intake (g), measured over a 24-h period, of control or PLX3397-treated mice given p.o. administration of either vehicle (H 2 O) or DON 12.5 ( B ) or 1.25 ( C ) mg/kg BW. A 2-way repeated-measures ANOVA ( p < 0.05) was performed in B and C followed by post hoc Bonferroni tests for individual time points. Differences in food intake between vehicle- and DON-treated groups was observed (** p < 0.01, *** p < 0.001). PLX3397 diet induced a significant difference in food intake of DON-treated groups ( ## p < 0.01, ### p < 0.001). D. Microphotographs illustrating c-Fos protein labeling observed with DVC (left), PVN (middle) and ARC (right) of control or PLX3397 mice treated with either vehicle or DON 1.25 mg/kg BW and killed 3 h after treatment. Scale bar: 200 µm. E. Quantification of c-Fos-positive cells in structures of interest. A 2-way repeated-measures ANOVA ( p < 0.05) was performed followed by post hoc Bonferroni tests for individual time points. Differences in c-Fos expression between vehicle- and DON-treated groups was observed (*** p < 0.001). The PLX3397 diet induced a significant difference between DON-treated groups ( ## p < 0.01, ### p < 0.001). AP area postrema; ARC arcuate nucleus; cc central canal; DVC dorsal vagal complex; ME median eminence; NTS nucleus tractus solitarius; PVN paraventricular nucleus; 3 V: third ventricle

Article Snippet: Counting of c-Fos positive nuclei was performed on photomicrographs acquired using a tenfold lens with a DXM 1200 Camera (Nikon) coupled to ACT-1 software.

Techniques: Staining, Labeling, Expressing

Journal: Journal of Neuroinflammation

Article Title: Constitutively active microglial populations limit anorexia induced by the food contaminant deoxynivalenol

doi: 10.1186/s12974-022-02631-7

Figure Lengend Snippet: Microglia inhibition by minocycline exacerbates DON-induced anorexia. A Representative images of CD68 staining in the DVC of animals treated with vehicle or minocycline. Scale bars: 200 µm. B Cumulative food intake (g) of control or minocycline-treated mice after receiving p.o. administration of vehicle (H 2 O) or DON 1.25 mg/kg BW for 24 h. C. Cumulative food intake (g), measured over a 6 h period, of control or minocycline-treated mice given having received p.o. or i.p. administration of DON 1.25 mg/kg BW. A 2-way repeated-measures ANOVA ( p < 0.05) was performed followed by Bonferroni post hoc tests for individual time points. *Significantly different from respective control group, * p < 0.05, ** p < 0.01, *** p < 0.001. # Significant difference between control and PLX3397 groups treated with DON; # p < 0.05; ### p < 0.001. D Microphotographs illustrating c-Fos protein labeling observed in DVC (left), PVN (middle) and ARC (right) of control or minocycline-treated mice that received either vehicle or DON 1.25 mg/kg BW and were killed 3 h after treatment. Scale bar: 200 µm. E Quantification of c-Fos positive cells in structures of interest for control and treated with DON mice. A 2-way repeated-measures ANOVA ( P < 0.05) was performed followed by Bonferroni post hoc tests for individual time points. Differences in c-Fos expression between vehicle- and DON-treated groups were observed (* p < 0.05; *** p < 0.001). PLX3397 diet induced a significant difference between DON-treated groups ( ### p < 0.001). AP area postrema; ARC arcuate nucleus; cc central canal; DVC dorsal vagal dorsal; fs funiculus separans ; ME median eminence; NTS nucleus tractus solitaries; PVN paraventricular nucleus; 3 V: third ventricle

Article Snippet: Counting of c-Fos positive nuclei was performed on photomicrographs acquired using a tenfold lens with a DXM 1200 Camera (Nikon) coupled to ACT-1 software.

Techniques: Inhibition, Staining, Labeling, Expressing