Journal: Brain, behavior, and immunity

Article Title: Chronic psychological stress and high-fat high-fructose diet disrupt metabolic and inflammatory gene networks in the brain, liver, and gut and promote behavioral deficits in mice

doi: 10.1016/j.bbi.2016.08.021

Figure Lengend Snippet: HFHF diet and stress impact the organization of tight junction proteins in the small intestine. Cryosections of small intestine were probed for ZO-1 (A) and CLDN2 (B) by immunostaining. HFHF diet and stress promote structural changes and disrupt the ZO-1 network in the small intestine (white arrows) (A). HFHF and stress increase CLDN2 lumenal surface localization in the small intestine (white arrows) (B). ZO-1 protein levels are unaffected by diet and stress (C), HFHF diet without stress significantly reduces abundance of OCLN protein (D); CLDN2 protein levels are unaffected by diet and stress, but ratio of high molecular weight to low molecular weight forms increases significantly with HFHF diet without stress (E). Images were obtained using a Nikon Eclipse 90i microscope with a DS-Fi1 (Nikon) camera and Nikon NIS-Elements AR 3.10 software. 40× mag - FITC 100 ms 1×) (CLDN2 40× mag – FITC 100 ms 1 ×). Protein expression in small intestine was measured by Western blot, band intensity calculated using densitometric analysis (Image Studio Lite), and values were normalized to β-actin. Data were analyzed by two-way ANOVA, with Bonferroni’s correction for multiple comparisons. Bar height indicates mean of samples from n = 6 mice per group, error bars indicate standard error of the mean (S.E.M.). Means with different letters are significantly different from each other (p<0.05). Abbreviations: ZO-1, Zonula occludens; CLDN2, Claudin-2; OCLN, Occludin; Control, Control diet; HFHF, High-fat high-fructose diet; NS, No stress, S, stress.

Article Snippet: Images were obtained using a Nikon Eclipse 90i microscope with a DS-Fi1 (Nikon) camera and Nikon NIS-Elements AR 3.10.

Techniques: Immunostaining, Molecular Weight, Microscopy, Software, Expressing, Western Blot

Journal: Brain, behavior, and immunity

Article Title: Chronic psychological stress and high-fat high-fructose diet disrupt metabolic and inflammatory gene networks in the brain, liver, and gut and promote behavioral deficits in mice

doi: 10.1016/j.bbi.2016.08.021

Figure Lengend Snippet: HFHF diet and stress modulate lipogenesis and inflammation in the liver. Gross morphologic changes in liver and gut from HFHF animals compared to the control groups (A). Light microscopic images of liver stained for lipids (B) reveal hepatocytes with normal appearance for the Control diet groups; and HFHF groups displaying lipid droplets (white arrows), cell infiltration (dashed area) and microvacuolization (black arrows). Oil Red O-stained and hematoxylin-counterstained liver sections, magnification 40×. Images were obtained using a Nikon Eclipse 90i microscope with a DS-Fi1 (Nikon) camera and Nikon NIS-Elements AR 3.10 software. HFHF S mice presented increased Lcn2 expression (C). HFHF diet decreases liver Tlr2 and Tlr4, (D, E) and stress reduces hepatic Tlr4 in the control diet group (E). Srebp1c RNA expression increases with HFHF treatment (F). There is no difference in hepatic Tnf mRNA expression (G). Bar height indicates mean of samples from an n = 6 mice per group, error bars indicate standard error of the mean (S.E.M.). Means with different letters are significantly different from each other (p < 0.05). Data were analyzed by two-way ANOVA followed by Bonferroni post hoc. Tissues were analyzed by qPCR using primers directed against murine Lcn2, Tlr2, Tlr4, Srebp1c, and Tnf. For each animal, the Ct values were normalized to the Ct values for Gapdh and Ppia. The relative expression level of the target gene (fold change) was expressed as 2−ΔΔCt, when compared with the mean DCt (threshold cycle) of the control group. Abbreviations: qPCR, quantitative real-time reverse-transcription polymerase chain reaction; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; TLR, Toll like receptor; SREBP-1c, sterol regulatory element binding protein-1c; Lcn2, neutrophil gelatinase-associated lipocalin 2; TNF, Tumor necrosis factor; Control, Control diet; HFHF, High-fat high-fructose diet; NS, No stress, S, stress.

Article Snippet: Images were obtained using a Nikon Eclipse 90i microscope with a DS-Fi1 (Nikon) camera and Nikon NIS-Elements AR 3.10.

Techniques: Staining, Microscopy, Software, Expressing, RNA Expression, Reverse Transcription Polymerase Chain Reaction, Binding Assay