Thermoneutral housing does not influence fat mass or glucose homeostasis in C57BL/6 mice

in Journal of Endocrinology
Correspondence should be addressed to A E Brandon: amanda.brandon@sydney.edu.au

*(L Small and H Gong contributed equally to this work)

One major factor affecting physiology often overlooked when comparing data from animal models and humans is the effect of ambient temperature. The majority of rodent housing is maintained at ~22°C, the thermoneutral temperature for lightly clothed humans. However, mice have a much higher thermoneutral temperature of ~30°C, consequently data collected at 22°C in mice could be influenced by animals being exposed to a chronic cold stress. The aim of this study was to investigate the effect of housing temperature on glucose homeostasis and energy metabolism of mice fed normal chow or a high-fat, obesogenic diet (HFD). Male C57BL/6J(Arc) mice were housed at standard temperature (22°C) or at thermoneutrality (29°C) and fed either chow or a 60% HFD for 13 weeks. The HFD increased fat mass and produced glucose intolerance as expected but this was not exacerbated in mice housed at thermoneutrality. Changing the ambient temperature, however, did alter energy expenditure, food intake, lipid content and glucose metabolism in skeletal muscle, liver and brown adipose tissue. Collectively, these findings demonstrate that mice regulate energy balance at different housing temperatures to maintain whole-body glucose tolerance and adiposity irrespective of the diet. Despite this, metabolic differences in individual tissues were apparent. In conclusion, dietary intervention in mice has a greater impact on adiposity and glucose metabolism than housing temperature although temperature is still a significant factor in regulating metabolic parameters in individual tissues.

 

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    Thermoneutrality reduces energy expenditure and energy intake without changing body or fat mass. 22°C = black. 29°C = red. Solid line/circles = chow. Dashed line/crosses = HFD. Mean ± s.e.m. (A) Weekly body mass. (B) Fat and (C) lean mass determined by echo MRI. n = 12. Analysed by repeated measures 2-way ANOVA for an effect of temperature and an effect of diet. (D) Diurnal oxygen consumption and (E) total over 24 h. (F) Diurnal respiratory exchange ratio and (G) average over 24 h. (H) Total energy intake over 24 h. (I) Total activity (beam breaks) over 24 h. n = 4. Analysed by 2-way ANOVA for an effect of temperature and an effect of diet. ***P < 0.0005. A full colour version of this figure is available at https://doi.org/10.1530/JOE-18-0279.

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    Effect of diet but no effect of temperature on glucose tolerance. 22°C = black. 29°C = red. Solid line/ circles = chow. Dashed line/crosses = HFD. Mean ± s.e.m. Oral glucose tolerance curves at (A) 8 and (E) 13 weeks from the start of study. Area under the curve of the glucose tolerance at (B) 8 and (F) 13 weeks. Blood insulin during the oGTT at (C) 8 and (G) 13 weeks. Area under the curve of the blood insulin at (D) 8 and (H) 13 weeks. Analysed by 2-way ANOVA for an effect of temperature and an effect of diet. n = 10–12. A full colour version of this figure is available at https://doi.org/10.1530/JOE-18-0279.

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    HFD-feeding increases UCP1 abundance in BAT while thermoneutrality reduces it. 22°C = black. 29°C = red. Mean ± s.e.m. (A) Representative immunoblot and (B) quantitation of UCP1 in BAT normalised to 14-3-3. n = 12 (C) 2-DG uptake (phosphorylated [3H]2-deoxyglucose) in BAT. n = 7–12. (D) Representative immunoblot and quantitation of subunits of mitochondrial OXPHOS Complex I-V in BAT, (E) Complex I (NDUFB8), (F) Complex II (SDHB), (G) Complex III (UQCRC2), (H) Complex IV (MTCO1) and (I) Complex V (ATP5A) normalised to 14-3-3. (J) BAT protein content. n = 10–12. Analysed by 2-way ANOVA for an effect of temperature and an effect of diet. ***P < 0.0005. (K) Hematoxylin & Eosin staining of BAT sections, scale bar = 25 µm. A full colour version of this figure is available at https://doi.org/10.1530/JOE-18-0279.

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    Increased triglyceride content and expression of lipogenic proteins in liver from mice at 29°C. 22°C = black. 29°C = red. Mean ± s.e.m. (A) Liver triglyceride content. (B) Liver glycogen content. (C) Representative immunoblots and quantitation of (D) ACC, (E) FAS, (F) SCD1 and (G) PEPCK normalised to 14-3-3. Analysed by 2-way ANOVA for an effect of temperature and an effect of diet. n = 11–12. *P < 0.05, **P < 0.005, ***P < 0.0005. (H) Hematoxylin & Eosin staining of liver sections, scale bar = 50 µm. A full colour version of this figure is available at https://doi.org/10.1530/JOE-18-0279.

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    Thermoneutrality reduces triglyceride content and increases basal glucose uptake in the quadriceps muscle of HFD mice. 22°C = black. 29°C = red. Mean ± s.e.m. (A) Quadriceps triglyceride content (B) Quadriceps β-HAD activity. n = 12. (C) 2-DG uptake (phosphorylated [3H]2-deoxyglucose) in the quadriceps. n = 7–12. (D) Quadriceps glycogen content. n = 11–12. (E) Linear regression of quadriceps triglyceride content and 2-DG uptake, n = 39, circles = chow, crosses = HFD, dashed lines = 95% confidence intervals. Representative immunoblot (F) and quantitation (G) of UCP3 in the quadriceps muscle normalised to β-Actin. n = 12. (E) Analysed by 2-way ANOVA for an effect of temperature and an effect of diet. **P < 0.005, ***P < 0.0005, #P = 0.051. A full colour version of this figure is available at https://doi.org/10.1530/JOE-18-0279.

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