No amyloid plaques were detected in transgenic mice using immunohistochemistry.

<p>Immunohistochemical staining of β-amyloid plaques using A) anti-Aβ (1–42) polyclonal antibody B) congo-red staining on cerebral sections of aging (10 month old) wild-type (WT, n = 3), ApoB-100 transgenic (TG, n = 3), and AD model (APPSwe×PSEN1dE9) mice (n = 3). Nuclei were stained using hematoxylin stain. Arrows indicate stained amyloid plaques. Scale bars represent 100 µm (A) and 200 µm (B).</p

Detection of increased triglyceride levels in the serum and cerebral tissues of ApoB-100 transgenic mice.

<p>A) Serum triglyceride levels of 3- and 9-month-old wild-type (WT, n = 6) and ApoB-100 transgenic (TG) mice (n = 6). Values represent the mean triglyceride level (mmol/l) ± SEM, ** denotes statistical significant difference of p<0.01. B) Detection of triglycerides in the brain of 10-month-old wild-type (WT) and ApoB-100 transgenic mice (TG) using Nile-red staining (n = 3). Scale bars denotes 100 µm. White arrow indicates accumulation of triglycerides as lipid droplet in the cortex of aging transgenic mice.</p

Increased expression of pTau Ser⁴⁰⁴ in transgenic animals.

<p>A) Representative picture of pTau Ser⁴⁰⁴ and amyloid precursor protein (APP) detection in young (3 month old) and adult (6-month-old) wild-type (WT) and ApoB-100 transgenic (TG) brain samples using western analysis B) Quantitative evaluation of western blots, (n = 3), ± SEM, * denotes statistically significant difference of p<0.05. C) Immunostaining of cortical and hippocampal regions from wild-type (WT) and ApoB-100 transgenic (TG) mice (n = 3) using anti-pTauSer⁴⁰⁴. Arrows indicate somatodendritic localization of hyperphosphorylated tau. HiF: hippocampal fissure; Mol: molecular layer of dentate gyrus. Scale bars represent 200 µm and 100 µm for the cortex and hippocampus, respectively.</p

Cortical lipoprotein profile of hyperlipidemic transgenic mice.

<p>A) Detection of major apolipoprotein and LDLr levels in the cortex of adult (6 month old) wild-type (WT) (n = 3) and ApoB-100 transgenic (TG) mice (n = 3) using western blotting. B) Quantification of western blot experiments. Relative OD values were expressed as % of wild-type mice (100%), ± SEM, * denotes a statistical difference of p<0.05.</p

Increased expression of ApoB-100 and ApoE proteins in the cortical and hippocampal tissues of ApoB-100 transgenic (TG) mice.

<p>Immunohistochemical detection of ApoB-100 in the cortex (upper row) and ApoE in the hippocampal region (lower row) of adult wild-type (WT) and ApoB-100 transgenic (TG) animals. Magnification 200×. Scale bars represent 100 µm.</p

Increased tau phosphorylation in the brain of adult (6 month old) ApoB-100 transgenic mice.

<p>A) Tau phosphorylation intensities were compared in the cortex of young (3 month-old) wild-type (WT, n = 3) and ApoB-100 transgenic (TG) mice (n = 3), using conventional, semi-quantitative western blottings. B) Quantification of western blots. First, Tau V-20 was normalized to the endogeneous β-actin, then different phosphosites to Tau V-20. C) Quantitative western analysis of adult (6 month old) wild-type (WT) and transgenic (TG) brain lysates. Phosphorylation level at different tau sites in wild-type mice represents 100%.</p

Impaired synaptic plasticity in hypertriglyceridemic mice.

<p>Electrophysiological recordings from brain slices of 3- and 6-month-old wild-type (Wt) and ApoB-100 transgenic (Tg) mice, (n = 6) A) Paired pulse facilitation (PPF), B) long-term synaptic plasticity. Arrows indicate theta burst stimulation (TBS) and low-frequency stimulation (LFS) C) detection of PKCγ expression in the brain of adult (6 month old) wild-type (WT) and transgenic mice (TG) using quantitative western blotting.</p

Altered Functional Protein Networks in the Prefrontal Cortex and Amygdala of Victims of Suicide

<div><p>Probing molecular brain mechanisms related to increased suicide risk is an important issue in biological psychiatry research. Gene expression studies on <em>post mortem</em> brains indicate extensive changes prior to a successful suicide attempt; however, proteomic studies are scarce. Thus, we performed a DIGE proteomic analysis of <em>post mortem</em> tissue samples from the prefrontal cortex and amygdala of suicide victims to identify protein changes and biomarker candidates of suicide. Among our matched spots we found 46 and 16 significant differences in the prefrontal cortex and amygdala, respectively; by using the industry standard <em>t</em> test and 1.3 fold change as cut off for significance. Because of the risk of false discoveries (FDR) in these data, we also made FDR adjustment by calculating the <em>q</em>-values for all the <em>t</em> tests performed and by using 0.06 and 0.4 as alpha thresholds we reduced the number of significant spots to 27 and 9 respectively. From these we identified 59 proteins in the cortex and 11 proteins in the amygdala. These proteins are related to biological functions and structures such as metabolism, the redox system, the cytoskeleton, synaptic function, and proteolysis. Thirteen of these proteins (CBR1, DPYSL2, EFHD2, FKBP4, GFAP, GLUL, HSPA8, NEFL, NEFM, PGAM1, PRDX6, SELENBP1 and VIM,) have already been suggested to be biomarkers of psychiatric disorders at protein or genome level. We also pointed out 9 proteins that changed in both the amygdala and the cortex, and from these, GFAP, INA, NEFL, NEFM and TUBA1 are interacting cytoskeletal proteins that have a functional connection to glutamate, GABA, and serotonin receptors. Moreover, ACTB, CTSD and GFAP displayed opposite changes in the two examined brain structures that might be a suitable characteristic for brain imaging studies. The opposite changes of ACTB, CTSD and GFAP in the two brain structures were validated by western blot analysis.</p> </div

Description of participants in the present study.

<p>NA: not available; S: suicide; C: control.</p

Western blot validation of GFAP, cathepsin and actin expressions in the cortex and amygdala of suicide and control subjects.

<p>The expressions of GFAP and cathepsin were significantly decreased in the suicide prefrontal cortex compared to the control samples while in the amygdala their expressions were significantly increased. In case of the actin similar but non-significant changes were found. The loading control was Ponceau, mean ± SEM.</p