Journal: Endocrinology

Article Title: A Mixed Glucocorticoid/Mineralocorticoid Selective Modulator With Dominant Antagonism in the Male Rat Brain.

doi: 10.1210/en.2015-1390

Figure Lengend Snippet: Figure 3. Fkbp5 and Sgk-1 expression in the hippocampus and dorsal striatum. A, C118335 blocked the corticosterone-induced up-regulation of Fkbp5 in the hippocampus. There was a significant effect of corticosterone treatment (F1,20 16.56; P .001) and of C118335 treatment (F1,20 16.23; P .001), and a significant corticosterone C118335 interaction (F1,20 7.301; P .05). B, C118335 down-regulated Sgk-1 expression in the hippocampus regardless of corticosterone treatment. A C118335 effect was observed (F1,20 25.71; P .001) and a marginally nonsignificant interaction effect (F1,20 4.127; P .06). C, No statistically significant C118335 effects were found on Fkbp5 expression in the striatum (F1,21 0.003; P .95). There was a trend toward an effect of corticosterone treatment (F1,21 3.274, P .084). D, C118335 treatment prevented the corticosterone-induced up-regulation of Sgk-1. There was a significant corticosterone treatment effect (F1,20 8.197; P .01) and a significant C118335 effect (F1,20 8.295; P .01). Bonferroni post hoc tests: *, P .05; ***, P .001; n 5 to 6 per group. E, Plasma corticosterone levels were high after vehicle treatment, but lower in C118335-treated animals (two-tailed t test: P .05). Arb. Units, arbitrary units.

Article Snippet: Spironolactone was purchased from Tocris (catalog number 2968) and cortisol and corticosterone were purchased from Sigma-Aldrich (catalog numbers H4001 and 27840, respectively).

Techniques: Expressing, Clinical Proteomics, Two Tailed Test

Journal: Endocrinology

Article Title: A Mixed Glucocorticoid/Mineralocorticoid Selective Modulator With Dominant Antagonism in the Male Rat Brain.

doi: 10.1210/en.2015-1390

Figure Lengend Snippet: Figure 4. Antagonism of C118335 in inhibitory avoidance. A, C118335 (80 mg/kg) administered subcutaneously 1 hour before inhibitory avoidance training antagonized the memory-enhancing effect of immediate posttraining corticosterone (1 mg/kg sc) treatment. Data show inhibitory avoidance retention latencies (mean SEM in seconds) on the 48-hour retention test. B, Immediate posttraining C118335 (50 or 80 mg/kg) induced dose-dependent impairment of memory consolidation of the inhibitory avoidance training that was mimicked by spironolactone (40 mg/kg sc) but not by RU486 (40 mg/kg sc). Data show inhibitory avoidance retention latencies (mean SEM in seconds) on the 48-hour retention test. **, P .01; ***, P .001; n 8 to 13 per group.

Article Snippet: Spironolactone was purchased from Tocris (catalog number 2968) and cortisol and corticosterone were purchased from Sigma-Aldrich (catalog numbers H4001 and 27840, respectively).

Techniques:

Journal: Nutrients

Article Title: Tryptophan Attenuates Chronic Restraint Stress-Induced Intestinal Injury Through Modulation of Intestinal Barrier Integrity and Gut Microbiota Homeostasis

doi: 10.3390/nu17060975

Figure Lengend Snippet: Figure 3. Tryptophan alleviates CRS-induced intestine leakage and epithelial apoptosis. (A) Serum intestinal fatty acid-binding protein content by ELISA; (B) FITC-dextran content in serum; (C) TUNEL staining plots; (D) TUNEL-positive cell counts; different letters denote significant differences between groups, p < 0.05. Ctrl refers to control group.

Article Snippet: We obtained the Mouse Corticosterone (CORT) ELISA Kit (Elabscience, Shanghai, China), FITC—dextran (4 kD) (Sigma, Saint Louis, MI, USA); the mouse intestinal fatty acid-binding protein (FABP) ELISA Kit (CUSABIO, Wuhan, China); and all of the 11 SCFA standards (ZZ Standards Co., Ltd., Shanghai, China).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Control

Journal: Nature Communications

Article Title: Psychological stress impairs IL22-driven protective gut mucosal immunity against colonising pathobionts

doi: 10.1038/s41467-021-26992-4

Figure Lengend Snippet: a Quantification of ileal Lcn2 by ELISA in control ( n = 16), stress ( n = 16), and LPS treated ( n = 8) mice. Significance was determined by a one-way ANOVA, not corrected for multiple comparisons, control:stress, p = 0.0141, control:LPS, p = 0.0466. b Ileal AIEC tissue burdens from control ( n = 16), stress ( n = 18), and LPS treated ( n = 10) mice control:stress, p < 0.0001, control:LPS, p = 0.0079. c Representative flow plots of CD45 + CD90 +/− cells and lineage staining on CD45 + CD90 + . Percentage of CD90 + T cells and CD90 + lineage − ILCs of total CD45 + cells isolated from ileal lamina propria cells collected from starved ( n = 10) and stressed ( n = 10) mice as determined by flow cytometry. Significance was determined by two-way ANOVA, not corrected for multiple comparisons, p < 0.0001, p < 0.0001. d Quantification of corticosterone in the serum as determined by ELISA in control ( n = 4), stress ( n = 6), and RU486 treated stress ( n = 6) mice. Significance was determined by one-way ANOVA, not corrected for multiple comparisons, control:stress, p = 0.0087; control:stress + RU486, p = 0.0011. e Representative flow plot of percentage of CD45 + CD90 + GR + cells. f Frequency of CD45 + CD90 + cells following 8 h of restraint stress in control ( n = 8), stress ( n = 12), and RU486 treated stress ( n = 11) mice. g Frequency of CD45 + CD90 + AnnexinV + cells following 8 h of restraint in control ( n = 8), stress ( n = 12), and RU486 treated stress ( n = 11). Significance was determined by a one-way ANOVA, not corrected for multiple comparisons, control:stress, p = 0.0323, stress:stress + ru486 p = 0.0018. h Frequency of CD45 + CD90 + cells following overnight restraint in control ( n = 18), stress ( n = 8), and RU486 treated stress ( n = 11). Significance was determined by one-way ANOVA, not corrected for multiple comparisons, control:stress, p = 0.0079, stress:stress + ru486 p = 0.0043. i Absolute number of CD45 + CD90 + cells following overnight restraint in control ( n = 18), stress ( n = 8), and RU486 treated stress ( n = 11) mice. Significance was determined by two-way ANOVA, not corrected for multiple comparisons, control:stress, p = 0.0005, stress:stress + ru486 p = 0.0060. (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001). Error bars represent SEM and the line in CFU graphs indicates the geometric mean of the group.

Article Snippet: Mouse corticosterone in the serum was enumerated using a Corticosterone Parameter Assay Kit (R&D Systems) as per the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Staining, Isolation, Flow Cytometry

Journal: International journal of molecular sciences

Article Title: Citrus Flavanone Effects on the Nrf2-Keap1/GSK3/NF-κB/NLRP3 Regulation and Corticotroph-Stress Hormone Loop in the Old Pituitary.

doi: 10.3390/ijms25168918

Figure Lengend Snippet: Figure 6. The effects of citrus flavanones on corticotroph cells in the pituitary, ACTH, and corti- costerone concentration of old rats. Immunohistochemical staining of ACTH (A), optical density of ACTH cells (B), plasma level of ACTH (C), and corticosterone level in serum (D). Asterisk (*) highlights dilated blood vessels with ACTH cells around them. 40× and 63× magnification, bar = 25 and 20 µm. Each value represents mean ± SD, n = 4; statistics: one-way ANOVA, Dunett’s multiple comparison post hoc test, * p < 0.05 citrus flavanone vs. CON rats. ICON, intact control; CON, control sunflower oil; NAR, naringenin; HES, hesperetin.

Article Snippet: Serum corticosterone concentrations were measured without dilution by immunoassay (KGE009, R&D Systems Inc., Minneapolis, MN, USA), in duplicate samples within a single assay, with an intra-assay CV of 8.0%.

Techniques: Concentration Assay, Immunohistochemical staining, Staining, Clinical Proteomics, Comparison, Control

Journal: Journal of internal medicine

Article Title: Circadian and ultradian rhythms: Clinical implications.

doi: 10.1111/joim.13795

Figure Lengend Snippet: Fig. 3 Transcriptional glucocorticoid (GC) receptor (GR)-dependent gene pulsing in rat liver and brain closely tracks the circulating GC ultradian rhythm. (a) Schematic depicting intracellular action of the GC hormones cortisol and corti- costerone, with cortisol shown here. Steroid hormones such as GCs are lipophilic and can pass through the lipid bilayer of the cell’s plasma membrane, where they act as ligands to bind and activate the intracellular GC receptors (GR) and mineralo- corticoid receptor (MR), with GR shown here. GR and MR are ligand-activated transcription factors. In the absence of ligand, the receptors are sequestered in the cytoplasm in complex with chaperone proteins including Heat Shock Protein 90 (HSP90), P23 and FK506 Binding Protein 51 (FKBP51). Upon ligand binding, the receptors undergo a conformational change allowing exchange of FKBP51 for FKBP52 and subsequent transport into the nucleus. Once in the nucleus, the ligand-bound receptor seeks out regulatory elements for GC target genes and initiates transcriptional induction of nascent messenger RNA, which is then exported back out to the cytoplasm and translated into protein by ribosomes. (c) Pulses of corticosterone delivered intravenously into adrenalectomized rats (denoted by P1, P2, P3) (red trace) induce transient hepatic GR activation (blue dashed trace) and pulses of Period 1 nascent RNA production in liver (green dashed trace) [52]. (c) Similarly, corticosterone pulses (light green trace) initiate transient hippocampal GR activation (blue dashed trace) and pulses of Period 1 nascent RNA production in rat brain (magenta dashed trace) [50]. Source: (a) Created with BioRender.com.

Article Snippet: Vehicle group = absence of hormone; CORT = 20 min corticosterone treatment (modelling a pulse peak); CORT + Wash = 40 min after the 20 min corticosterone treatment is removed from cells by stringent washing in a hormone-free cell medium (modelling a pulse nadir). (C) University of California Santa Cruz Genome Browser image at the Fkbp5 gene locus shows the comparison of mapped MR and GR DNA binding sites for each treatment group described in (B).

Techniques: Clinical Proteomics, Membrane, Binding Assay, Ligand Binding Assay, Activation Assay

Journal: Journal of internal medicine

Article Title: Circadian and ultradian rhythms: Clinical implications.

doi: 10.1111/joim.13795

Figure Lengend Snippet: Fig. 4 Pulse phase determines response to stress. (a) Corticosterone profiles of rats subjected to an auditory stress during automated blood sampling (LHS panel) revealed a greater corticosterone response in rats that were stressed during the ris- ing phase compared to the falling phase of endogenous corticosterone pulses. The effect was revealed in post hoc analysis that separated the data into two groups, with the stressor coinciding with either the rising or falling phase of an endogenous corticosterone pulse. The dotted lines indicate the noise stress interval. (b) A causal relation was demonstrated in adrenalec- tomized rats infused with pulsatile corticosterone for 12 h before an auditory stress was delivered at defined times, minutes into either the rising or falling phases of the corticosterone pulse. The ACTH response was greater in the rising phase than the falling phase. Neuronal activation in the amygdala was greater when the stress was encountered during the rising than the falling phase. Behavioural analysis revealed an overall increase in specific risk assessment behaviour activity in the rats subjected to the noise stress during the rising phase compared to the falling phase.

Article Snippet: Vehicle group = absence of hormone; CORT = 20 min corticosterone treatment (modelling a pulse peak); CORT + Wash = 40 min after the 20 min corticosterone treatment is removed from cells by stringent washing in a hormone-free cell medium (modelling a pulse nadir). (C) University of California Santa Cruz Genome Browser image at the Fkbp5 gene locus shows the comparison of mapped MR and GR DNA binding sites for each treatment group described in (B).

Techniques: Sampling, Activation Assay, Activity Assay

Journal: Journal of internal medicine

Article Title: Circadian and ultradian rhythms: Clinical implications.

doi: 10.1111/joim.13795

Figure Lengend Snippet: Fig. 5 Dynamics of glucocorticoid (GC) receptor (GR) and mineralocorticoid receptor (MR) during ultradian GC exposure. (A) Hippocampal GR and MR activation dynamics in response to two intravenous corticosterone (CORT) pulses administered to adrenalectomized rats (representative plot adapted from [46]). (B) Venn diagrams show the proportions of GR and MR DNA binding sites that directly overlap by at least 1 bp between treatments (i) vehicle, (ii) CORT pulse, and (iii) washout period after CORT pulse [71]. Vehicle group = absence of hormone; CORT = 20 min corticosterone treatment (modelling a pulse peak); CORT + Wash = 40 min after the 20 min corticosterone treatment is removed from cells by stringent washing in a hormone-free cell medium (modelling a pulse nadir). (C) University of California Santa Cruz Genome Browser image at the Fkbp5 gene locus shows the comparison of mapped MR and GR DNA binding sites for each treatment group described in (B). This track shows an estimate of the binding activity of mapped to genomic coordinates presented in a Genome Browser. The Genome Browser annotation tracks page displays the precise genome location of sites of GR or MR binding to the DNA, as specified through an uploaded custom annotation track containing the enriched short DNA sequences that had been directly bound by the transcription factor of interest, in this case GR and MR.

Article Snippet: Vehicle group = absence of hormone; CORT = 20 min corticosterone treatment (modelling a pulse peak); CORT + Wash = 40 min after the 20 min corticosterone treatment is removed from cells by stringent washing in a hormone-free cell medium (modelling a pulse nadir). (C) University of California Santa Cruz Genome Browser image at the Fkbp5 gene locus shows the comparison of mapped MR and GR DNA binding sites for each treatment group described in (B).

Techniques: Activation Assay, Binding Assay, Comparison, Activity Assay

Journal: Journal of internal medicine

Article Title: Circadian and ultradian rhythms: Clinical implications.

doi: 10.1111/joim.13795

Figure Lengend Snippet: Fig. 6 Experimental design for the novel experimental shift work model in rats. (a) Corticosterone (CORT) profiles are mon- itored in blood samples taken every 10 min using an automated blood sampler (ABS). A programmable syringe pump is used to infuse adrenalectomized rats with circadian and ultradian patterned corticosterone at physiologically levels, either in-phase (b) or out-of-phase (c) with light–dark cues.

Article Snippet: Vehicle group = absence of hormone; CORT = 20 min corticosterone treatment (modelling a pulse peak); CORT + Wash = 40 min after the 20 min corticosterone treatment is removed from cells by stringent washing in a hormone-free cell medium (modelling a pulse nadir). (C) University of California Santa Cruz Genome Browser image at the Fkbp5 gene locus shows the comparison of mapped MR and GR DNA binding sites for each treatment group described in (B).

Techniques: