Ed food intake. (C) MIC-1/GDF15-treated MIC-12/2 and (D) MIC-

Ed food intake. (C) MIC-1/GDF15-treated MIC-12/2 and (D) MIC-1/GDF15-treated MIC-1+/+ consumed 23388095 significantly less food than the matched vehicle-treated mice of same genotype (MIC-12/2 n = 6/group, p = 0.04; MIC-1+/+ n = 14/group, p,0.01 unpaired t-test). Data expressed as mean 6 SE. doi:10.1371/journal.pone.0055174.AN 3199 manufacturer gMIC-1/GDF15 Regulates Appetite and Body Weightnormalized to bodyweight compared to the age matched control MIC-1+/+ mice (p,0.01, Fig. 5B, 5D). This difference may be partially attributed to a decrease in physical activity, since physical activity was significantly decreased during the dark phase 1531364 in female MIC-12/2 versus control mice (p = 0.03, Fig. 5C, 5E). No such differences in energy expenditure or physical activity were observed between MIC-12/2 and MIC-1+/+ male mice (Fig. 4B, 4C, 4D, 4E). To determine the likely contribution of changes in physical activity to changes in energy expenditure, correlation analysis was performed using hourly data from individual mice. There was a positive correlation between energy expenditure and physical activity within all the groups (p,0.02 by Pearson correlation for all groups, Fig. 6A and 6B). In both males and females, the difference in the slope of the regression line is significantly different for MIC12/2 and MIC-1+/+ mice (p,0.01 in all group, Fig. 6), indicating that the energy cost of activity was different between genotypes. Further, to estimate basal metabolic rate, the function from the trend line was used to extrapolate physical activity to zero, with the point at which the line crosses the X-axis signifying basal metabolic rate. While there was no genotypic difference in basal metabolic rate between males (Fig. 6A), female MIC-12/2 displayed a significantly lower basal metabolic rate compared to wild type controls (p,0.01) (Fig. 6B). These data indicate that there is a fundamental metabolic difference between the male and female MIC-12/2 mice relatively to their matched controls, suggesting MIC-1/GDF15 exerts its effects differentially on male and female mice. The difference in total energy expenditure between the female genotypic groups may have been more LY-2409021 affected by changes in basal metabolic rate and less by the physical activity. Both of these are likely to contribute to the body weight difference displayed in female MIC-12/2 versus control mice.MIC-1/GDF15 Reduces Food Intake and Induces Weight Loss in MIC-12/2 and MIC-1+/+ MiceTo investigate whether the increased body weight of MIC-12/2 mice was specifically due to germline gene deletion of MIC-1/ GDF15 and not resulting from any unrecognized compensatory or developmental changes in MIC-12/2 mice, we continuously infused male MIC-12/2 and MIC+/+ mice with MIC-1/GDF15 (1? mg/d) via osmotic minipumps. Infusion of MIC-1/GDF15 over 5 days resulted in increased circulating human MIC-1/ GDF15 levels from zero to 643667 pg/ml and 576645 pg/ml in MIC-12/2 and MIC-1+/+ mice, respectively. As discussed below, this would have the effect of increasing total MIC-1/GDF15 levels in MIC-12/2 mice to about the middle of the human normal range and in MIC-1+/+ mice to the top of the human normal range. This infusion of MIC-1/GDF15 resulted in reduced body weight gain relative to syngeneic vehicle-infused controls (Fig. 7A and 7B, MIC-12/2 p,0.01; MIC-1+/+ p = 0.01), coupled with a significant reduction in food intake (Fig. 7C and 7D, MIC-12/2 p = 0.04; MIC-1+/+ p,0.01).DiscussionIn addition to high circulating levels of MIC-1/GDF15 me.Ed food intake. (C) MIC-1/GDF15-treated MIC-12/2 and (D) MIC-1/GDF15-treated MIC-1+/+ consumed 23388095 significantly less food than the matched vehicle-treated mice of same genotype (MIC-12/2 n = 6/group, p = 0.04; MIC-1+/+ n = 14/group, p,0.01 unpaired t-test). Data expressed as mean 6 SE. doi:10.1371/journal.pone.0055174.gMIC-1/GDF15 Regulates Appetite and Body Weightnormalized to bodyweight compared to the age matched control MIC-1+/+ mice (p,0.01, Fig. 5B, 5D). This difference may be partially attributed to a decrease in physical activity, since physical activity was significantly decreased during the dark phase 1531364 in female MIC-12/2 versus control mice (p = 0.03, Fig. 5C, 5E). No such differences in energy expenditure or physical activity were observed between MIC-12/2 and MIC-1+/+ male mice (Fig. 4B, 4C, 4D, 4E). To determine the likely contribution of changes in physical activity to changes in energy expenditure, correlation analysis was performed using hourly data from individual mice. There was a positive correlation between energy expenditure and physical activity within all the groups (p,0.02 by Pearson correlation for all groups, Fig. 6A and 6B). In both males and females, the difference in the slope of the regression line is significantly different for MIC12/2 and MIC-1+/+ mice (p,0.01 in all group, Fig. 6), indicating that the energy cost of activity was different between genotypes. Further, to estimate basal metabolic rate, the function from the trend line was used to extrapolate physical activity to zero, with the point at which the line crosses the X-axis signifying basal metabolic rate. While there was no genotypic difference in basal metabolic rate between males (Fig. 6A), female MIC-12/2 displayed a significantly lower basal metabolic rate compared to wild type controls (p,0.01) (Fig. 6B). These data indicate that there is a fundamental metabolic difference between the male and female MIC-12/2 mice relatively to their matched controls, suggesting MIC-1/GDF15 exerts its effects differentially on male and female mice. The difference in total energy expenditure between the female genotypic groups may have been more affected by changes in basal metabolic rate and less by the physical activity. Both of these are likely to contribute to the body weight difference displayed in female MIC-12/2 versus control mice.MIC-1/GDF15 Reduces Food Intake and Induces Weight Loss in MIC-12/2 and MIC-1+/+ MiceTo investigate whether the increased body weight of MIC-12/2 mice was specifically due to germline gene deletion of MIC-1/ GDF15 and not resulting from any unrecognized compensatory or developmental changes in MIC-12/2 mice, we continuously infused male MIC-12/2 and MIC+/+ mice with MIC-1/GDF15 (1? mg/d) via osmotic minipumps. Infusion of MIC-1/GDF15 over 5 days resulted in increased circulating human MIC-1/ GDF15 levels from zero to 643667 pg/ml and 576645 pg/ml in MIC-12/2 and MIC-1+/+ mice, respectively. As discussed below, this would have the effect of increasing total MIC-1/GDF15 levels in MIC-12/2 mice to about the middle of the human normal range and in MIC-1+/+ mice to the top of the human normal range. This infusion of MIC-1/GDF15 resulted in reduced body weight gain relative to syngeneic vehicle-infused controls (Fig. 7A and 7B, MIC-12/2 p,0.01; MIC-1+/+ p = 0.01), coupled with a significant reduction in food intake (Fig. 7C and 7D, MIC-12/2 p = 0.04; MIC-1+/+ p,0.01).DiscussionIn addition to high circulating levels of MIC-1/GDF15 me.