Induction of Salivary Proteins Modifies Measures of Both Orosensory and Postingestive Feedback during Exposure to a Tannic Acid Diet - Figure 4

<p>A–D: White bars represent feeding behaviors measured while animals were consuming the control diet; grey bars represent the same behaviors measured while animals were on the 3% tannic acid diet. The white bar labeled ‘C’ represents a 5-day average of behavioral measures on the control diet prior to exposure to the tannic acid diet. Food intake (A) and meal size (B) were decreased on the first 3 days of exposure to the tannic acid diet but returned to control levels by day 4. Meal number (C) was increased during the first 2 days of exposure to the tannic acid diet but returned to control-levels by day 3. Rate of feeding (D) was decreased throughout the entire exposure to the tannic acid diet buy this effect was most pronounced during the first 3 days.</p

Establishing Variations in Adiposity in Diet-Induced Obesity in Male C57BL/6J Mice

<p>Frequency distribution of body weight in 219 mice at 12 wk of age, after being fed a high-fat diet for 4 wk. (A) Regression analysis between the change in body weight and fat mass (B) and change in body weight and lean body mass (C) from 8 and 12 wk of age as determined by NMR in 112 mice. (D) Regression analysis between mouse weight at weaning and the percent change of the ratio of fat mass (FM) to lean mass (LM) per week, <i>p</i> = .0105, <i>n</i> = 220. Changes in the FM/LM ratio per wk were calculated from 8–12 wk of age or from 8–14 wk of age. (E) Average percent change of the ratio of FM/LM per week for the various litter sizes, <i>p</i> = .0406, <i>n</i> = 220 (ANOVA).</p

Over-Expression of Four Genes in High Gainer Mice in Two Peritoneal Fat Depots and One Subcutaneous Fat Depot Illustrates that the Mechanism Leading to Over-Expression of These Genes Occurs in All Fat Depots

<p>Statistical significance was calculated by ANOVA. Twenty mice were present in each group.</p

Data depicted in the larger graphs are densitometry units normalized to average control (water replete) protein expressions (which are set to 1).

<p>The first bar (stippled) represents the average water replete expression of protein concentration, while all rats were fed the control diet. The data in the remainder of the bars were collected in the water-deprived condition. The white bar represents the average protein expression of all rats on the day of their first exposure to the brief-access taste test. The white bar with hash marks represents the saliva samples collected the day of the second exposure to the brief-access test by rats that were maintained on the control diet. The gray bar with hash marks represents the saliva samples collected the day of the second exposure to the brief-access test by rats that were maintained on the tannic acid diet between exposures. Statistical analyses were not preformed on these data. Total protein concentration was significantly altered by water deprivation making comparisons between samples unreliable. We have presented them only to illustrate the relative abundance of proteins across treatments. The inset graphs represent the change in densitometry units between the two test sessions (i.e. test 1 protein expression- test 2 protein expression) for the control group (white bar with hash marks) and experimental group (gray bar with hash marks). *Experimental group greater than the control group, P<0.05.</p

Coordinated Gene Expression

<p>Regression analyses of SFRP5, MEST, and Naked mRNA levels in RNA isolated from high and low gainer mice in inguinal fat depots suggest that regulation of this subset of genes share a common mechanism related to the degree of adiposity. Twenty mice were present in each group.</p

The sequence of rat protein cystatin S (Uniprot accession P19313).

<p>Bold regions indicate where tryptic peptides detected in proteomic analysis overlay the protein sequence.</p

Summary of linear mixed model analyses.

<p>We looked for the predictive value of each of the normalized densitometry units of protein bands at each of the listed masses (kDa) on feeding behaviors. Analyses were restricted to the first four days of exposure to the tannin diet as this represented the dynamic phase over which changes in feeding behavior and salivary protein expression were observed. *ps<0.05.</p><p>Summary of linear mixed model analyses.</p

Positive Correlations between SFRP5 and Adiposity

<div><p>Regression analysis between SFRP5 mRNA levels and adiposity as estimated by the ratio of FM to LM in ING, EPI, and RP fat from 112 Mice Described in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.0020081#pgen-0020081-g001" target="_blank">Figure 1</a>B.</p><p>FM = Fat Mass; LM = Lean Mass; ING = Inguinal; EPI = Epididymal; RP = Retroperitoneal</p></div

Representative samples of the glycosylation determination.

<p>The gel contains saliva samples from a single rat after 4 or 12 days exposure to the tannic acid diet. The samples were loaded in duplicate on the same12% SDS/PAGE. The quantity of protein loaded in each lane was varied to allow for maximal visibility of the protein band. After resolving the gel, it was cut into two pieces. Gel A was stained using Pro-Q Emerald 300 glycoprotein detection kit, which detects only glycosylated proteins. Gel B was stained with Coomassie R250 stain, which detects total proteins. Both gels contain both Candy-Cane glycoprotein molecular weight standards and Bio-Rad molecular weight standards.</p

Summary of ANOVAs comparing normalized densitometry units of protein bands at each of the listed masses (kDa).

<p>Analyses were conducted with experimental group (control or tannic acid diet) as the between-subjects comparison and time (days) as the within-subjects comparison. *ps<0.05.</p><p>Summary of ANOVAs comparing normalized densitometry units of protein bands at each of the listed masses (kDa).</p