Consumption of a 13-week HFS diet at 20 weeks of age did not exacerbate age-related declines in bone or muscle, but these tissues do not decline in a coordinate manner with greater declines in bone than muscle.

Contraction‐mediated lipolysis increases the association of lipid droplets and mitochondria, indicating an important role in the passage of fatty acids from lipid droplets to mitochondria in skeletal muscle. PLIN3 and PLIN5 are of particular interest to the lipid droplet–mitochondria interaction because PLIN3 is able to move about within cells and PLIN5 associates with skeletal muscle mitochondria. This study primarily investigated: 1) if PLIN3 is detected in skeletal muscle mitochondrial fraction; and 2) if mitochondrial protein content of PLIN3 and/or PLIN5 changes following stimulated contraction. A secondary aim was to determine if PLIN3 and PLIN5 associate and whether this changes following contraction. Male Long Evans rats (n = 21; age, 52 days; weight = 317 ± 6 g) underwent 30 min of hindlimb stimulation (10 msec impulses, 100 Hz/3 sec at 10–20 V; train duration 100 msec). Contraction induced a ~50% reduction in intramuscular lipid content measured by oil red‐O staining of red gastrocnemius muscle. Mitochondria were isolated from red gastrocnemius muscle by differential centrifugation and proteins were detected by western blotting. Mitochondrial PLIN5 content was ~1.6‐fold higher following 30 min of contraction and PLIN3 content was detected in the mitochondrial fraction, and unchanged following contraction. An association between PLIN3 and PLIN5 was observed and remained unaltered following contraction. PLIN5 may play a role in mitochondria during lipolysis, which is consistent with a role in facilitating/regulating mitochondrial fatty acid oxidation. PLIN3 and PLIN5 may be working together on the lipid droplet and mitochondria during contraction‐induced lipolysis.

Calcineurin is a Ca2+/calmodulin (CaM)-dependent phosphatase that plays a critical role in promoting the slow fiber phenotype and myoblast fusion in skeletal muscle, thereby making calcineurin an attractive cellular target for enhancing fatigue resistance, muscle metabolism, and muscle repair. Neurogranin (Ng) is a CaM-binding protein thought to be expressed solely in brain and neurons, where it inhibits calcineurin signaling by sequestering CaM, thus lowering its cellular availability. Here, we demonstrate for the first time the expression of Ng protein and mRNA in mammalian skeletal muscle. Both protein and mRNA levels are greater in slow-oxidative compared with fast-glycolytic muscles. Coimmunoprecipitation of CaM with Ng in homogenates of C2C12 myotubes, mouse soleus, and human vastus lateralis suggests that these proteins physically interact. To determine whether Ng inhibits calcineurin signaling in muscle, we used Ng siRNA with C2C12 myotubes to reduce Ng protein levels by 60%. As a result of reduced Ng expression, C2C12 myotubes had enhanced CaM-calcineurin binding and calcineurin signaling as indicated by reduced phosphorylation of nuclear factor of activated T cells and increased utrophin mRNA. In addition, calcineurin signaling affects the expression of myogenin and stabilin-2, which are involved in myogenic differentiation and myoblast fusion, respectively. Here, we found that both myogenin and stabilin-2 were significantly elevated by Ng siRNA in C2C12 cells, concomitantly with an increased fusion index. Taken together, these results demonstrate the expression of Ng in mammalian skeletal muscle where it appears to be a novel regulator of calcineurin signaling.

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While repeated in vivo micro-computed tomography (μCT) allows for longitudinal measurement of bone outcomes in rodent models, it is important to determine that the resulting irradiation – dependent on the frequency and number of scans - does not exceed the effects of the intervention. The objective of this study was to determine whether repeated irradiation exposure from μCT scans at 1-month intervals for a total of four scans would alter trabecular or cortical bone structure outcomes and/or bone mineral density in tibias from both male and female CD-1 mice. The right tibia of male (n = 12) and female (n = 11) CD-1 mice were scanned using μCT at 2, 3, 4, and 5 months of age, while the contralateral left tibia served as a control and was scanned only at 5 months of age. All scans were performed at a resolution of 9 μm using a radiation dose of 460 mGy per scan. Some outcomes of trabecular bone structure were affected by repeated irradiation in both males and females. The bone volume fraction was lower in the irradiated right tibia compared to the non-irradiated left tibia in both males (p < 0.05) and females (p < 0.01) as a result of decreased trabecular number (males p < 0.05; females p < 0.05) and increased trabecular separation (males p < 0.05; females p < 0.01). Some cortical measures were also affected in females but not in males, including lower cortical bone periosteal perimeter (p < 0.05), lower total area (p < 0.01) and lower marrow area (p < 0.05) with repeated irradiation. Exposure to repeated radiation at intervals of 1 month, for a total of four scans, altered trabecular bone in both male and female CD-1 mice while outcomes of cortical bone structure were altered only in females.