Characterization of sclerostin’s response within white adipose tissue to an obesogenic diet at rest and in response to acute exercise in male mice

This study examined the effect of a high-fat diet (HFD) on sclerostin content within subcutaneous inguinal visceral white adipose tissue (iWAT), and visceral epididymal WAT (eWAT) depots at rest and in response to acute aerobic exercise. Male C57BL/6 mice (n=40, 18 weeks of age) underwent 10 weeks of either a low-fat diet (LFD) or HFD. Within each diet group, mice were assigned to either remain sedentary (SED) or perform 2h of endurance treadmill exercise at 15 m·min-1 with 5° incline (EX), creating 4 groups: LFD+SED (N=10), LFD+EX (N=10), HFD+SED (N=10), and HFD+EX (N=10). Serum and WAT depots were collected 2h post-exercise. Serum sclerostin showed a diet-by-exercise interaction, reflecting HFD+EX mice having higher concentration than HFD-SED (+31%, p=0.03), and LFD mice being unresponsive to exercise. iWAT sclerostin content decreased post-exercise in both 28 kDa (-31%, p=0.04) and 30 kDa bands (-36%, main effect for exercise, p=0.02). iWAT b-catenin (+44%, p=0.03) and GSK3b content were elevated in HFD mice compared to LFD (+128%, main effect for diet, p=0.005). Monomeric sclerostin content was abolished in eWAT of HFD mice (-96%, main effect for diet, p<0.0001), was only detectable as a 30 kDa band in LFD mice and was unresponsive to exercise. b-catenin and GSK3b were both unresponsive to diet and exercise within eWAT. These results characterized sclerostin’s mobilization to WAT depots in response to acute exercise, which appears to be specific to a reduction in iWAT and identified a differential regulation of sclerostin’s form/post-translational modifications depending on diet and WAT depot.This research was funded by the Natural Sciences and Engineering Research Council of Canada (NSERC grant to P. Klentrou # 2020-00014). N. Kurgan, B. Baranowski and Joshua Stoikos hold NSERC doctoral scholarships

Bone Turnover Markers and Osteokines in Adolescent Female Athletes of High- and Low-Impact Sports Compared With Nonathletic Controls

This study examined differences in resting concentrations of markers of bone formation and resorption, and osteokines between female adolescent (12–16 y) swimmers, soccer players, and nonathletic controls. Resting, morning blood samples were obtained after an overnight fast from 20 swimmers, 20 soccer players, and 20 nonathletic controls, matched for age. carboxyl-terminal cross-linking telopeptide of type I collagen (CTX), amino-terminal propeptide of type I collagen (P1NP), total osteocalcin (OC), sclerostin, osteoprotegerin (OPG), and receptor activator of nuclear factor kappa B ligand (RANKL) were analyzed in serum. After controlling for percent body fat, there were no significant differences between swimmers and nonathletic controls in any of the measured markers. In contrast, soccer players had significantly higher P1NP (89.5 [25.6] ng·mL−1), OC (57.6 [22.9] ng·mL−1), and OPG (1052.5 [612.6] pg·mL−1) compared with both swimmers (P1NP: 66.5 [20.9] ng·mL−1; OC: 24.9 [12.5] ng·mL−1; OPG: 275.2 [83.8] pg·mL−1) and controls (P1NP: 58.5 [16.2] ng·mL−1; OC: 23.2 [11.9] ng·mL−1; OPG: 265.4 [97.6] pg·mL−1), with no differences in CTX, sclerostin, and RANKL. These results suggest that bone formation is higher in adolescent females engaged in high-impact sports like soccer compared with swimmers and controls

A Low-Therapeutic Dose of Lithium Inhibits GSK3 and Enhances Myoblast Fusion in C2C12 Cells

Glycogen synthase kinase 3 (GSK3) slows myogenic differentiation and myoblast fusion partly by inhibiting the Wnt/β-catenin signaling pathway. Lithium, a common medication for bipolar disorder, inhibits GSK3 via Mg+ competition and increased Ser21 (GSK3α) or Ser9 (GSK3β) phosphorylation, leading to enhanced myoblast fusion and myogenic differentiation. However, previous studies demonstrating the effect of lithium on GSK3 have used concentrations up to 10 mM, which greatly exceeds concentrations measured in the serum of patients being treated for bipolar disorder (0.5–1.2 mM). Here, we determined whether a low-therapeutic (0.5 mM) dose of lithium could promote myoblast fusion and myogenic differentiation in C2C12 cells. C2C12 myotubes differentiated for three days in media containing 0.5 mM lithium chloride (LiCl) had significantly higher GSK3β (ser9) and GSK3α (ser21) phosphorylation compared with control myotubes differentiated in the same media without LiCl (+2–2.5 fold, p < 0.05), a result associated with an increase in total β-catenin. To further demonstrate that 0.5 mM LiCl inhibited GSK3 activity, we also developed a novel GSK3-specific activity assay. Using this enzyme-linked spectrophotometric assay, we showed that 0.5 mM LiCl-treated myotubes had significantly reduced GSK3 activity (−86%, p < 0.001). Correspondingly, 0.5 mM LiCl treated myotubes had a higher myoblast fusion index compared with control (p < 0.001) and significantly higher levels of markers of myogenesis (myogenin, +3-fold, p < 0.001) and myogenic differentiation (myosin heavy chain, +10-fold, p < 0.001). These results indicate that a low-therapeutic dose of LiCl is sufficient to promote myoblast fusion and myogenic differentiation in muscle cells, which has implications for the treatment of several myopathic conditionsBrock University Library Open Access Publishing Fun

The Effect of Sclerostin on Myotropic Response to Exercise

This study is the first to examine the effect of exogenous sclerostin injection on myotropic responses to sedentary behaviour and an aerobic exercise intervention. Specifically, changes in myosin heavy chain isoform distribution, muscle mass and myofiber cross sectional area were studied in response to sedentary behaviour and to a 5-week aerobic exercise intervention. Mice (n=24) were assigned to either remain sedentary (SED, n=24), or assigned to a 4-week exercise training program (EXT, n=24) and further spit into their final groupings with sedentary control (SED+C) and exercise control (EXT+C) groups receiving saline injections and sedentary sclerostin (SED+S) and exercise sclerostin (EXT+S) groups recombinant sclerostin. Soleus and extensor digitorum longus muscle were then extracted and analyzed via fluorescent immunohistochemistry and Western blotting. Sclerostin injection led to a significant reduction in soleus MHCI, MHC1/IIA, MHCIIA/X and MHC IIB cross-sectional area (p < .5) along with trending declines in MHC IIA (p = .1). In contrast to this, there was no effect of sclerostin injection on MHC IIX or MHCIIXB CSA. In EDL tissue there was a trend towards a decrease in tissue necropsy weight in the sclerostin injection group (p = .1). Sclerostin appeared to have no effect on total MHC protein content or in the examined markers of Wnt signaling (GSK3β, β-catenin) as detected in both soleus and EDL muscle tissue via western blot. Our findings demonstrate that sclerostin negatively influences muscle tissue via decreases in myofibril cross-sectional area and these decreases trend towards a significant reduction in muscle mass

DataSheet2_Characterization of sclerostin’s response within white adipose tissue to an obesogenic diet at rest and in response to acute exercise in male mice.PDF

Introduction: It is well established that sclerostin antagonizes the anabolic Wnt signalling pathway in bone, however, its physiological role in other tissues remains less clear. This study examined the effect of a high-fat diet (HFD) on sclerostin content and downstream markers of the Wnt signaling pathway (GSK3β and β-catenin) within subcutaneous inguinal white adipose tissue (iWAT), and visceral epididymal WAT (eWAT) depots at rest and in response to acute aerobic exercise.Methods: Male C57BL/6 mice (n = 40, 18 weeks of age) underwent 10 weeks of either a low-fat diet (LFD) or HFD. Within each diet group, mice were assigned to either remain sedentary (SED) or perform 2 h of endurance treadmill exercise at 15 m min−1 with 5° incline (EX), creating four groups: LFD + SED (N = 10), LFD + EX (N = 10), HFD + SED (N = 10), and HFD + EX (N = 10). Serum and WAT depots were collected 2 h post-exercise.Results: Serum sclerostin showed a diet-by-exercise interaction, reflecting HFD + EX mice having higher concentration than HFD + SED (+31%, p = 0.03), and LFD mice being unresponsive to exercise. iWAT sclerostin content decreased post-exercise in both 28 kDa (−31%, p = 0.04) and 30 kDa bands (−36%, main effect for exercise, p = 0.02). iWAT β-catenin (+44%, p = 0.03) and GSK3β content were higher in HFD mice compared to LFD (+128%, main effect for diet, p = 0.005). Monomeric sclerostin content was abolished in eWAT of HFD mice (−96%, main effect for diet, p Conclusion: These results characterized sclerostin’s content to WAT depots in response to acute exercise, which appears to be specific to a reduction in iWAT and identified a differential regulation of sclerostin’s form/post-translational modifications depending on diet and WAT depot.</p

DataSheet1_Characterization of sclerostin’s response within white adipose tissue to an obesogenic diet at rest and in response to acute exercise in male mice.PDF

Introduction: It is well established that sclerostin antagonizes the anabolic Wnt signalling pathway in bone, however, its physiological role in other tissues remains less clear. This study examined the effect of a high-fat diet (HFD) on sclerostin content and downstream markers of the Wnt signaling pathway (GSK3β and β-catenin) within subcutaneous inguinal white adipose tissue (iWAT), and visceral epididymal WAT (eWAT) depots at rest and in response to acute aerobic exercise.Methods: Male C57BL/6 mice (n = 40, 18 weeks of age) underwent 10 weeks of either a low-fat diet (LFD) or HFD. Within each diet group, mice were assigned to either remain sedentary (SED) or perform 2 h of endurance treadmill exercise at 15 m min−1 with 5° incline (EX), creating four groups: LFD + SED (N = 10), LFD + EX (N = 10), HFD + SED (N = 10), and HFD + EX (N = 10). Serum and WAT depots were collected 2 h post-exercise.Results: Serum sclerostin showed a diet-by-exercise interaction, reflecting HFD + EX mice having higher concentration than HFD + SED (+31%, p = 0.03), and LFD mice being unresponsive to exercise. iWAT sclerostin content decreased post-exercise in both 28 kDa (−31%, p = 0.04) and 30 kDa bands (−36%, main effect for exercise, p = 0.02). iWAT β-catenin (+44%, p = 0.03) and GSK3β content were higher in HFD mice compared to LFD (+128%, main effect for diet, p = 0.005). Monomeric sclerostin content was abolished in eWAT of HFD mice (−96%, main effect for diet, p Conclusion: These results characterized sclerostin’s content to WAT depots in response to acute exercise, which appears to be specific to a reduction in iWAT and identified a differential regulation of sclerostin’s form/post-translational modifications depending on diet and WAT depot.</p