1. Volume 15, Issue 2, Pages 171-185 (February 2012)
Altered Mitochondrial Function and Metabolic Inflexibility Associated with Loss of Caveolin-1 
Ingrid Wernstedt Asterholm, Dorothy I. Mundy, Jian Weng, Richard G.W. Anderson, Philipp E. Scherer 
Cell Metabolism 
Volume 15, Issue 2, Pages (February 2012)
DOI: /j.cmet
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2. Cell Metabolism 2012 15, 171-185DOI: (10.1016/j.cmet.2012.01.004)
Copyright © 2012 Elsevier Inc. Terms and Conditions

3. Figure 1
Caveolin-1 Null Mice Display Reduced Circulating Adiponectin Levels and Reduced Sensitivity to β3-AR Agonist
(A) mRNA and protein levels of adiponectin in 3-month-old male mice (n = 7–11 per group).
(B) Circulating levels of adiponectin in 2-month-old mice (n = 10–12 per group).
(C) Adiponectin complex distribution in male mice matched for total levels of adiponectin (n = 3 per group).
(D and E) FFA and insulin response to β3-AR agonist (1 mg/kg i.p.) in 2-month-old male mice (n = 4–5 per group). The error bars indicate SEM.
Cell Metabolism  , DOI: ( /j.cmet )
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4. Figure 2
Reduced Hepatic Steatosis, Reduced Response to ADF, Lower RER Variability, but Normal Levels of Fasting-Induced FFAs in the Caveolin-1 Null Mice
Hepatic steatosis was assessed by CT measurements before and after a 24 hr fast (A) (note that the difference is plotted). (B) Representative ORO stain of livers from 24 hr fasted mice. Hepatic steatosis was assessed after 8 weeks of HFD feeding ad libitum (C) and followed by 4 weeks HFD feeding ADF or ad libitum HFD (D). Chow-fed mice were acclimatized to the metabolic cages for 4 days, and on the fifth day RER (E) was recorded. Male mice 3–4 months old were used for (A)–(E) (n = 4–10 per group). (F) and (G) show the levels of circulating FFAs and glycerol in the fed state at 7 p.m.; 24 hr fasted state at 7 p.m.; and 15, 30, and 60 min after refeeding (n = 5 per group; 12-week-old female cohorts used). The error bars indicate SEM.
Cell Metabolism  , DOI: ( /j.cmet )
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5. Figure 3
Caveolin-1 Null Mice Have Increased Fasting Glucose Levels, Increased Breakdown of Amino Acids, and an Enhanced Response to Phospodiesterase Inhibitor
(A and B) Shown are the levels of circulating insulin and glucose in the fed state at 7 p.m.; 24 hr fasted state at 7 p.m.; and 15, 30, and 60 min after refeeding.
(C) Glycogen levels in 5 hr fasted male mice (12 weeks old; n = 5 per group).
(D) Weight loss after 24 hr fasting.
(E and F) Circulating urea (E) and circulating as well as hepatic acetyl-carnitine levels (F) were measured in 5 hr male fasted mice (3 months old; n = 7–9 per group). Enoximone (10 mg/kg i.p.) was injected to 5 hr fasted male mice (4 months old; n = 3–5 per group).
(G and H) Enoximone-induced changes in glucose and glycerol levels. The error bars indicate SEM.
Cell Metabolism  , DOI: ( /j.cmet )
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6. Figure 4
Caveolin-1 Null Mice Display an Altered Response to the PEPCK Inhibition
Acute response to PEPCK inhibition: 12-week-old female cohorts were fasted for 5 hr at daytime prior to intragastric administration of 30 mg/kg 3-MPA (n = 5 per group). Glucose levels were transiently lowered (A), while lactate (B) and glycerol (C) increased in the caveolin-1 null mice. Chronic response to PEPCK inhibition was as follows: fasting-induced hepatic steatosis, followed by CT in female mice (8 months old; n = 5 per group), was aggravated in the presence of PEPCK inhibitor (D). Glucose levels were measured at the end of the time course and remained higher in both PBS- and 3-MPA-treated caveolin-1 null mice (E). (F) Glycerol-induced glucose production measured in PEPCK-inhibited female mice (3 months old; n = 4–5 per group) was higher in caveolin-1 nulls, indicating that the higher glucose levels in caveolin-1 null mice during fasting are mediated by an increase in glycerol-induced gluconeogenesis. The error bars indicate SEM.
Cell Metabolism  , DOI: ( /j.cmet )
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7. Figure 5
Caveolin-1 Deficiency Results in Reduced Hepatic Triglyceride Synthesis, Increase in Reactive Oxygen Species, Elevated Adipose Tissue FFAs, and Increased Macrophage Infiltration
(A) Intravenous infusion of 3H-triolein in mice fasted for 5 hr reveals a reduced hepatic triglyceride synthesis in the liver of caveolin-1 null mice. Note that the 3H count in the organic phase of adipose tissue (BAT and IWAT) was increased in the caveolin-1 null mice. This indicates an increase in triglyceride synthesis rate.
(B and C) Circulating H2O2 and pyruvate levels in 2-month-old mice after 5 hr of fasting (n = 10–12 per group).
(D) Adiponectin and pyruvate levels measured in 4-month-old female mice were positively correlated.
(E) Local FFA levels from gonadal adipose tissue pieces (n = 4 + 4) obtained from 5-month-old females.
(F and G) Gonadal adipose tissue stained for mac2 and perilipin, respectively. The error bars indicate SEM.
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8. Figure 6 Altered Mitochondrial Function in Cav-1 Null Cells
(A) Caveolin-1−/− MEFs display a preference for glycolysis. Cells were incubated in various concentrations (in mM) of pyruvate (P) and glucose (G) as indicated. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured in a Seahorse XF flux analyzer.
(B) Caveolin-1 null cells have a higher mitochondrial membrane potential as judged by TMRM fluorescence intensity.
(C) Comparison of wild-type and caveolin-1 null dicarboxylate carrier (mDIC) mRNA levels in MEFs, adipose tissue, and liver. All measurements reflect triplicate measurements.
(D) BCAA levels after 5 hr of fasting in 2-month-old mice (n = 10–12 per group).
(E) Phospho-mTOR levels were measured in m. gastrocnemius in 5 hr fasted, 3-month-old male mice (n = 6 per group).
(F) Shown is schematic representation of caveolin-1 null phenotype: caveolin-1 deficiency leads to a reduced sensitivity to insulin and adrenergic agonists and elevated basal lipolysis, and is associated with altered mitochondrial function in adipose tissue. These defects have a profound impact on whole-body metabolism. In the caveolin-1 null mice, we observe a dramatically enhanced hepatic glucose production as well as an increase in BCAA levels (likely due to increased food intake). We hypothesize that the elevated lipolysis and the altered mitochondrial function in adipose tissue (and possibly other tissues) expose the liver to elevated levels of gluconeogenic substrates. The elevated BCAA levels both enhance fatty acid oxidation and increase anabolic processes in skeletal muscle. The error bars indicate SEM.
Cell Metabolism  , DOI: ( /j.cmet )
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