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Isolation of extracellular fluids reveals novel secreted bioactive proteins from muscle and fat tissues

ABSTRACT: Proteins are secreted from cells to send information to neighboring cells or distant tissues. Because of the highly integrated nature of energy balance systems, there has been particular interest in myokines and adipokines. These are challenging to study through proteomics because serum or plasma contain highly abundant proteins that limit the detection of proteins with lower abundance. We show here that extracellular fluid (EF) from muscle and fat tissues of mice show a different protein composition than either serum or tissues. Mass spectrometry analyses of EFs from mice with physiological perturbations, like exercise or cold exposure, allowed quantification of many potentially novel myokines and adipokines. Using this approach, we identify prosaposin as a secreted product of muscle and fat. Prosaposin expression stimulates thermogenic gene expression and induces mitochondrial respiration in primary fat cells. These studies together illustrate the utility of EF isolation as a discovery tool for adipokines and myokines.

ORGANISM(S): Mus musculus

PROVIDER: GSE216094 | GEO | 2023/01/20

REPOSITORIES: GEO

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Project description:Calorie restriction (CR) and exercise training (EX) are two critical lifestyles for the prevention and treatment of metabolic diseases, such as obesity and diabetes. Brown adipose tissue (BAT) and skeletal muscle (SkM) are two important organs to generate heat. Here we provide detailed transcriptional profiling of these two thermogenic tissues from mice treated with CR and/or EX. We demonstrate the transcriptional reprogramming of BAT and SkM according to CR, but little to EX. Consistent with this, CR induces genes encoding adipokines or myokines alteration in BAT and SkM, respectively. Deconvolution analysis implies differences of subpopulations of myogenic cells, mesothelial cells and endogenic cells in BAT, and demonstrates differences of subpopulations of satellite cells, immune cells and endothelial cells in SkM according to CR or EX. NicheNet analysis, exploring potential inter-organ communication, indicates that BAT and SkM can mutually regulate their fatty acid metabolism and thermogenesis by ligands and receptors. These data comprise an extensive resource for thermogenic tissues molecular responses to CR and/or EX in healthy state. Calorie restriction (CR) and exercise training (EX) are two critical lifestyles for the prevention and treatment of metabolic diseases, such as obesity and diabetes. Brown adipose tissue (BAT) and skeletal muscle (SkM) are two important organs to generate heat. Here we provide detailed transcriptional profiling of these two thermogenic tissues from mice treated with CR and/or EX. We demonstrate the transcriptional reprogramming of BAT and SkM according to CR, but little to EX. Consistent with this, CR induces genes encoding adipokines or myokines alteration in BAT and SkM, respectively. Deconvolution analysis implies differences of subpopulations of myogenic cells, mesothelial cells and endogenic cells in BAT, and demonstrates differences of subpopulations of satellite cells, immune cells and endothelial cells in SkM according to CR or EX. NicheNet analysis, exploring potential inter-organ communication, indicates that BAT and SkM can mutually regulate their fatty acid metabolism and thermogenesis by ligands and receptors. These data comprise an extensive resource for thermogenic tissues molecular responses to CR and/or EX in healthy state.

Project description:The underlying mechanism of how the atopic lipids in skeletal muscle affect muscle growth remains elusive. Here we chose miniature Bama swine as our model to mimick human obesity and co-associated metabolic disorders by long time diet induction and study how the atopic fat accumulation in skeletal muscle influence muscle function. After 23 months high-fat high-sucrose diet (HFHSD) fed, the model minipig model of obesity accompanied with metabolic disorders like human, and they had increased body weight and extensive lipids deposition in adipose tissues (AT) and non-AT, especially in skeletal muscle. Further more, the mass of skeletal reduced greatly and the small area (0-2000μm2) muscle reduced after diet induced. The average fiber area of Gastroc reduced 25.2%, but no significant changes appeared in the other skeletal muscles. Antioxidant capacity of skeletal muscle also reduced. Microarray profiles showed genes related to fat deposition promotion (Peroxisome proliferator activated receptor γ, CCAAT/enhancer-binding protein α and apolipoprotein E), muscle growth inhibition (myostatin and p21) up regulated, and some other muscle cell differentiation related genes (myoD) down regulated. Meanwhile, adipokines like adiponectin and 11b-hydroxysteroid dehydrogenase type 1 (11βHSD1) which partake in the crosstalk between AT and skeletal muscles rose up. We draw a clear potential crosstalk pathway that, increased 11βHSD1 secreted by excess AT will promote the expression of the major inhibitor MSTN by activating corticosterone to cortisol, leading to the growth inhibition of skeletal muscle. Overall, this research announces how obesity affects skeletal muscle growth in a crosstalk sight. Male and female Bama minipigs, aged 6 months at the start of the study, were divided into the following two groups for 23 months of treatment. Bama minipigon control (CD group, N=3) were fed standard pig chow. The experimental group (N=6) were fed high-fat high-sucrose diet (53% basal diet, 37% sucrose, 10% lard, HFHSD).

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Isolation of extracellular fluids reveals novel secreted bioactive proteins from muscle and fat tissues

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