Examples of POMC, NPY and GAD₆₅ mRNA Expression Following Olanzapine Treatment.

<p>Examples of pro-opiomelanocortin (POMC), neuropeptide Y (NPY) and glutamic acid decarboxylase 65 (GAD₆₅) mRNA expression in the female Sprague Dawley rat brain following 2.0 mg/kg olanzapine treatment (<b>A-A′, C-C′, E-E′</b>) or vehicle (control) (<b>B-B′, D-D′, F-F′</b>) for 14-days (t.i.d.). (<b>A–F</b>): Low magnification film autoradiographs depicting mRNA expression in the rat brain, (<b>A′–F′</b>): High magnification emulsion/cresyl violet–stained slides showing mRNA expression specifically in the arcuate nucleus. Autoradiographs are examples of raw data used for the graphs depicted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033548#pone-0033548-g003" target="_blank">Figure 3A–C</a> and are average representations of 6 rats per treatment group.</p

Weight Gain and Visceral Adiposity.

<p>(<b>A</b>): Percentage body weight change and (<b>B</b>): visceral adiposity in female Sprague Dawley rats (<i>n</i> = 12/treatment group) treated with 0.25, 0.5, 1.0 or 2.0 mg/kg olanzapine or vehicle (control), (14-days, t.i.d). Data expressed as mean±SEM. *<i>p</i><0.05, **<i>p</i><0.01 vs. control.</p

Correlations.

<p>Correlations between pro-opiomelanocortin (POMC) mRNA expression in the arcuate (Arc) nucleus and (<b>A</b>): percentage of body weight change, (<b>B</b>): visceral adipose tissue, (<b>C</b>): neuropeptide Y (NPY) mRNA expression in the Arc, (<b>D</b>): glutamic acid decarboxylase (GAD₆₅) mRNA expression in the Arc, (<b>E</b>): Arc NPY and GAD₆₅ mRNA expression, (<b>F</b>): cannabinoid CB1 receptor (CB1R) binding density in the dorsal vagal complex (DVC) and percentage of body weight changed, (<b>G</b>): DVC CB1R binding density and visceral adipose tissue, and (<b>H</b>): DVC CB1R binding density and GAD₆₅ mRNA expression, following 14-days olanzapine treatment or vehicle (control). Correlation analyses were made from raw data underlying the graphs presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033548#pone-0033548-g001" target="_blank">Figures 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033548#pone-0033548-g003" target="_blank">3</a>. Key: <>\raster="rg1"<> control, ×0.5 mg/kg, ○ 0.5 mg/kg, □ 1.0 mg/kg, ▵ 2.0 mg/kg olanzapine.</p

Example of CB1R Binding Density Following Olanzapine Treatment.

<p>Example of cannabinoid CB1 receptor binding density (using [³H]SR-141716A) in the <b>A, C:</b> hypothalamic arcuate nucleus and <b>B, D:</b> dorsal vagal complex of the caudal brainstem of female Sprague Dawley rats following <b>A, B:</b> 2.0 mg/kg olanzapine treatment, <b>C, D:</b> vehicle (control) for 14-days (t.i.d.). Autoradiographs are examples of raw data used for the graphs depicted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033548#pone-0033548-g003" target="_blank">Figure 3D</a> and are average representations of 6 rats per treatment group.</p

Dosage Effects of Olanzapine Treatment on POMC, NPY and GAD₆₅ mRNA Expression, and CB1R Binding Density.

<p>(<b>A</b>): Pro-opiomelanocortin mRNA expression (nCi/g tissue), (<b>B</b>): neuropeptide Y mRNA expression (nCi/g tissue), (<b>C</b>): glutamic acid decarboxylase (GAD₆₅) mRNA expression (nCi/g tissue), (<b>D</b>): cannabinoid CB1 receptor binding density (fmoles/mg tissue), in rats treated with 0.25. 0.5, 1.0 or 2.0 mg/kg olanzapine or vehicle (control) (14-days, t.i.d.) (<i>n</i> = 6/treatment group). Key: ▪ arcuate nucleus □ dorsal vagal complex. Data is expressed as mean ± SEM. *<i>p</i><0.05, **<i>p</i><0.01 vs. control.</p

Mean fat mass, liver weight, and body length (mean ± SEM) in female Sprague Dawley rats treated with olanzapine (1 mg/kg, t.i.d.) and/or betahistine (9.6 mg/kg, t.i.d.) or control (vehicle).

<p>*, <i>p</i><0.05;</p><p>**, <i>p</i><0.01 <i>vs.</i> control;</p>#<p>, <i>p</i><0.05;</p>##<p>, <i>p</i><0.01 <i>vs.</i> olanzapine.</p

Effects of olanzapine and/or betahistine treatment (n = 12) on lipid droplet deposition of hepatic tissue.

<p>A–C: HE staining of hepatic tissue from rats treated with Vehicle (A), Olanzapine-only (B), Betahistine-only (C), and O+B co-treatment (D). E: Fat cell counts on the liver sections of different treatment groups. F: Total fat cell area measured on the liver sections of different treatment groups. ** <i>p</i><0.01 <i>vs.</i> control; ## <i>p</i><0.01 <i>vs.</i> olanzapine.</p

Effects of olanzapine and/or betahistine treatment on body weight gain.

<p>A: Outline of the experimental design. B: The trend of three phases of drug administration on the accumulated body weight side-effect. Olanzapine (1 mg/kg, t.i.d.; n = 12), betahistine (9.6 mg/kg, t.i.d.; n = 12), co-treatment (O+B; n = 12) or control (vehicle; n = 12) for 11 weeks. (♦: control •: olanzapine, x: betahistine, ▴: O+B co-treatment). * <i>p</i><0.05, ** <i>p</i><0.01 <i>vs.</i> control, # <i>p</i><0.05 <i>vs.</i> olanzapine.</p

Effects of olanzapine and/or betahistine treatment on the hypothalamic protein levels of histamine H₁R, AMPKα, pAMPKα, and POMC.

<p>A: Examples of the images of the western blot experiment showing the protein expressions of histamine H₁R, AMPKα, pAMPKα, POMC and β-actin (n = 6). B–F: Effects of olanzapine and/or betahistine treatment on protein expressions of (B) hypothalamic H₁R, (C) AMPKα, (D) pAMPKα, (E) POMC, (F) neuropeptide Y (NPY). Abbreviations: H₁R: H₁ receptor, AMPKα: AMPK-activated protein kinase α, pAMPKα: the AMPK phosphorylation α and POMC: proopiomelanocortin. * <i>p</i><0.05, ** <i>p</i><0.01 <i>vs.</i> control; # <i>p</i><0.05, # <i>p</i><0.05 <i>vs.</i> olanzapine.</p

Effects of olanzapine and/or betahistine treatment on food intake and feeding efficiency.

<p>A–B: Accumulated food intake (A) and feeding efficiency (B) in the first phase of olanzapine treatment (1 mg/kg, t.i.d.; n = 12) compared with vehicles. C–D: Food intake (C) and Feeding efficiency (D) following olanzapine withdrawal. E–F: Food intake (E) and feeding efficiency (F) following chronic treatment of olanzapine (1 mg/kg, t.i.d.; n = 12), betahistine (9.6 mg/kg, t.i.d.; n = 12), co-treatment (O+B; n = 12) or vehicle (control; n = 12) for 5 weeks. (♦: control •: olanzapine, x: betahistine, ▴: O+B co-treatment). * <i>p</i><0.05, ** <i>p</i><0.01, olanzapine <i>vs.</i> control; $<i>p</i><0.05,$$ <i>p</i><0.01, co-treatment of O+B <i>vs.</i> control; # <i>p</i><0.05 O+B <i>vs.</i> olanzapine.</p