11β-HSD1 content in visceral fat is attenuated with RU486 treatment but not with C113176 or C108297 treatment.

<p>No changes were found in lipolytic protein levels with treatment of antagonists. 11β-HSD1 content was measured in epididymal fat pads to represent visceral adipose tissue and expressed relative to α-tubulin content (A). CD36 protein levels were measured from epididymal fat pads (B) as well as adipose triglyceride lipase (ATGL) (C) and hormone-sensitive lipase (HSL) (D) protein levels as markers of lipolytic adipose tissue activity and expressed relative to loading control. Bars that do not share similar letters denote statistical significance, p<0.05 one-way ANOVA using Tukey's post-hoc test. n = 5–6. All values are means ± SE.</p

Plasma CORT concentration.

<p>Plasma CORT concentration.</p

Glucose stimulated insulin secretion (GSIS) was normalized with RU486 and C113176 treatment.

<p>GSIS in isolated islets was measured in low (2.8 mM) and high (16.7 mM) glucose media for 1-hour incubations expressed as ng/ml/islet/hour. Bars that do not share similar letters denote statistical significance, p<0.05 one-way ANOVA using student's unpaired t-test. n = 3–7. All values are means ± SE.</p

Corticosterone-induced capillary rarefaction is abrogated by continuous prazosin treatment.

<p>TA muscle were sectioned and stained for capillaries using fluorescein<i>-Griffonia Simplicifolia</i> isolectin and Cy3-anti-α smooth muscle actin. (A) Representative images of isolectin staining after 1W and 2W of CORT-treatment. Inverted grey scale images of isolectin staining are displayed to enhance visualization of individual muscle fibers. Scale bar represents 100 μm. (B) C:F at the 1W and (C) 2W time points was calculated from the average of 5 non-overlapping fields of view, ^#<i>P</i><0.05 1W CORT vs. corresponding control group; ***, ^###<i>P</i><0.001 2W CORT vs. corresponding water or control group respectively, n = 6–9.</p

C113176 treatment maintains body mass while RU486 increases body mass with ROD treatment.

<p>Animal body mass (g) were recorded every two days for 10 days as a measure of fold change from day 0, pellet surgery (A). Animal body mass on day 10 was measured as a percent change of body mass from day 0 (B). The dotted line (100%) represents no change in body mass from day 0. Arrow indicates that 2 days after pellet surgeries respective antagonists or vehicle were administered at 80 mg/kg/day to each treatment group. Bars that do not share similar letters denote statistical significance, p<0.05, one-way ANOVA using Tukey's post-hoc test. n = 7–10. All values are means ± SE.</p

Fat accumulation is normalized with RU486 in skeletal muscle and liver cross sections.

<p>To determine fat content in skeletal muscle, tibialis anterior muscle was dissected and stained with a neutral lipid stain (Oil Red O) (A–E). Cross sections of liver were also stained with Oil Red O to measure lipid content (F–J).</p

Glucose intolerance and acute insulin response (AIR) is improved with RU486 and C113176 treatment.

<p>Fasting (basal, 0 minutes) and stimulated blood glucose levels (mM) were measured at 5, 15, 30, 60, 90 and 120 minutes post oral glucose gavage (A). Glucose area under the curve (AUC) was calculated based on fasting blood glucose of individual animals (A′). Fasting (basal, 0 minutes) and glucose-stimulated insulin levels (ng/ml) were measured at 15, 30, 60, and 120 minutes post oral glucose gavage (B). Insulin area under the curve (AUC) was calculated based on fasting individual insulin levels within each group (B′). To measure insulin capacity acute insulin response (AIR) was measured by the difference in insulin levels between fasting insulin and 15 minutes post glucose gavage (C). Negative values represent a decrease in insulin response, indicating impairment in insulin secretion. Bars that do not share similar letters denote statistical significance, p<0.05, one-way ANOVA using Tukey's post-hoc. A student's unpaired t-test was performed between controls and ROD, C108297 and C113176 groups (C). n = 7–10. All values are means ± SE.</p

Alterations to VEGF-A and TSP-1 with elevated CORT and concomitant prazosin treatment.

<p>RNA or protein was isolated from the TA muscle after 1W or 2W of CORT with or without concurrent prazosin treatment. Taqman qPCR was used to assess the mRNA levels of VEGF-A (A,B) and TSP-1 (D,E), while VEGF-A protein was assessed by ELISA (C) and TSP-1 protein by Western blot (F). (A) VEGF-A mRNA was not altered in response to 1W CORT and/or prazosin. (B) VEGF-A mRNA was not affected by 2W CORT (<i>P</i> = 0.08), while a significant prazosin effect was detected within the 2W CORT-prazosin group (*<i>P</i><0.05). (C) At the 2W time point, VEGF-A protein displayed a tendency for a CORT effect (<i>P</i> = 0.08), and was significantly increased in the CORT-prazosin cohort compared to water CORT animals (*<i>P</i><0.05). (D, E) TSP-1 mRNA was not altered by 1W or 2W CORT and/or prazosin. (F) TSP-1 protein level was significantly reduced with 2W CORT treatment (^#<i>P</i><0.05), and there was a trend (<i>P</i> = 0.09) for an increase in TSP-1 after 2W of prazosin treatment within control animals (n = 4–9).</p

Corticosterone concentrations, absolute and relative food intake.

<p>Note: BM = Body Mass. Different letters denote statistical significance, p<0.05, n = 6–10. The * indicates that a significant difference was performed by a student's unpaired t-test. All values are mean ± SE.</p

Influence of corticosterone on endothelial specific shear stress responsiveness.

<p>Cultured rat skeletal muscle endothelial cells were pre-treated with corticosterone (600 nM) for 48 hours prior to shear stress (15dynes/cm²), or were maintained in static conditions (C), for 2 hours. (A-D): Whole cell lysates were analyzed by Western blotting. (A) Phospho-ERK1,2 protein level relative to β-actin. (B) Phospho-p38 protein level relative to total p38. (C) Phospho-Ser473 Akt protein level relative to total Akt. (D) Phospho-Thr308 Akt protein level relative to total Akt. Two-way ANOVA indicated a significant shear effect for all kinases (pERK1,2: <i>P</i> = 0.007; pp38: <i>P</i> = 0.02; pSerAkt: <i>P</i> = 0.02; pThrAkt: <i>P</i> = 0.0003, n = 3–4). *,** <i>P</i><0.05, <i>P</i><0.01 compared to respective static control, post hoc analysis. A significant interaction between shear stress and CORT was detected only for pAktSer473 (^#<i>P</i> = 0.05). (E) Nitric oxide level was assessed indirectly by Griess assay. A main effect of shear stress was detected (<i>P</i> = 0.0006; n = 6). (*,**<i>P</i><0.05 and <i>P</i><0.01, relative to static controls as assessed via post hoc analysis).</p