RNA-sequencing reveals a gene expression signature in skeletal muscle of a mouse model of age-associated post-operative functional decline

2021 ◽  
Author(s):  
Samantha L Asche-Godin ◽  
Zachary A Graham ◽  
Adina Israel ◽  
Lauren M Harlow ◽  
Weihua Huang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Mouse Model ◽  
Physical Function ◽  
Grip Strength ◽  
Rna Sequencing ◽  
Gastrocnemius Muscle ◽  
Motor Coordination ◽  
Surgical Stress ◽  
Forelimb Grip Strength

This study aimed to characterize the effects of laparotomy on post-operative physical function and skeletal muscle gene expression in C57BL/6N mice at 3, 20 and 24 months of age to investigate late-life vulnerability and resiliency to acute surgical stress. Pre- and post-operative physical functioning were assessed by forelimb grip strength and motor coordination. Laparotomy induced an age-associated post-operative decline in forelimb grip strength that was greatest in the oldest mice. In contrast, while motor coordination declined with increasing age at baseline, it was unaffected by laparotomy. Moreover, baseline physical function as stratified by motor coordination performance (low vs. high functioning) in 24-month-old mice did not differentially affect post-laparotomy reduction in grip strength. RNA sequencing of soleus muscles showed that laparotomy induced age-associated differential gene expression and canonical pathway activation with the greatest effects in the youngest mice. Examples of such age-associated, metabolically important pathways that were only activated in the youngest mice after laparotomy included oxidative phosphorylation and NRF2-mediated oxidative stress response. Analysis of lipid mediators in serum and gastrocnemius muscle showed alterations in profiles of these mediators during aging and confirmed an association between such changes and functional status in gastrocnemius muscle. These findings demonstrate a mouse model of laparotomy which recapitulated some features of post-operative skeletal muscle decline in older adults following surgery, and identified age-associated, laparotomy-induced molecular signatures in skeletal muscles. Future research can build upon this mouse model to study molecular mechanisms of late-life vulnerability to acute surgical stress and resiliency to counter surgery-induced physical decline.

scholarly journals EPA and DHA Inhibit Myogenesis and Downregulate the Expression of Muscle-related Genes in C2C12 Myoblasts

Genes ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 64 ◽  
Author(s):  
Jing Zhang ◽  
Xin Xu ◽  
Yan Liu ◽  
Lin Zhang ◽  
Jack Odle ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Rna Sequencing ◽  
Cell Counting ◽  
Sequencing Analysis ◽  
C2c12 Myoblasts ◽  
Epa And Dha ◽  
Proliferation And Differentiation

This study was conducted to elucidate the biological effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on cell proliferation, differentiation and gene expression in C2C12 myoblasts. C2C12 were treated with various concentrations of EPA or DHA under proliferation and differentiation conditions. Cell viability was analyzed using cell counting kit-8 assays (CCK-8). The Edu assays were performed to analyze cell proliferation. To analyze cell differentiation, the expressions of myogenic marker genes were determined at the transcriptional and translational levels by qRT-PCR, immunoblotting and immunofluorescence. Global gene expression patterns were characterized using RNA-sequencing. Phosphorylation levels of ERK and Akt were examined by immunoblotting. Cell viability and proliferation was significantly inhibited after incubation with EPA (50 and 100 μM) or DHA (100 μM). Both EPA and DHA suppressed C2C12 myoblasts differentiation. RNA-sequencing analysis revealed that some muscle-related genes were significantly downregulated following EPA or DHA (50 μM) treatment, including insulin-like growth factor 2 (IGF-2), troponin T3 (Tnnt3), myoglobin (Mb), myosin light chain phosphorylatable fast skeletal muscle (Mylpf) and myosin heavy polypeptide 3 (Myh3). IGF-2 was crucial for the growth and differentiation of skeletal muscle and could activate the PI3K/Akt and the MAPK/ERK cascade. We found that EPA and DHA (50 μM) decreased the phosphorylation levels of ERK1/2 and Akt in C2C12 myoblasts. Thus, this study suggested that EPA and DHA exerted an inhibitory effect on myoblast proliferation and differentiation and downregulated muscle-related genes expression.

scholarly journals Modified forelimb grip strength test detects aging-associated physiological decline in skeletal muscle function in male mice

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Hikari Takeshita ◽  
Koichi Yamamoto ◽  
Satoko Nozato ◽  
Tadakatsu Inagaki ◽  
Hirotsugu Tsuchimochi ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Grip Strength ◽  
Muscle Function ◽  
Strength Test ◽  
Male Mice ◽  
Skeletal Muscle Function ◽  
Forelimb Grip Strength

scholarly journals PO-184 Effects of resistance training and aerobic training on Fibronectin of Skeletal Muscle Extracellular Matrix and Satellite Cell in Aging Mice

2018 ◽  
Vol 1 (4) ◽  
Author(s):  
Jian Lu ◽  
Shanshan Li ◽  
Caizhen Chen
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Matrix ◽  
Resistance Training ◽  
Protein Content ◽  
Grip Strength ◽  
Satellite Cell ◽  
Gastrocnemius Muscle ◽  
Aerobic Training ◽  
Significant Difference ◽  
Group A

Objective To investigate the effects of resistance and aerobic training on the content of fibronectin in skeletal muscle extracellular matrix and satellite cell in 9-month-old BALB/c mice, and then explore the effect of fibronectin content change on satellite cell and its intrinsic mechanism, which will provide a new research perspective and theoretical evidence for delaying sarcopenia. Methods 27 one-month-old BALB / c mice of SPF grade were purchased and fed for natural aging. At 9 months of age, the mice were divided into three groups randomly. Group R was resistance exercise training group (n=9), group A was aerobic exercise training group (n=9), and group C was control group (n=9). The load ladder model for resistance training in group R, the pyramid training program for 9 week training, 5 sets of load ladder training every time. 2 min for rest between sets, 1 min for rest between repetitions, 3 times a week. The treadmill training for aerobic training in group A for 9 weeks, the speed of 0.8km/h, 40min every time, 3 times a week. There is no training in group C. During the training, mouse grip strength was tested by the BIOSEB grip instrument once a week. After exercise intervention, the blood of mice was taken from the eyeball, and the gastrocnemius muscles were removed and placed in -80℃ temperature refrigerator to be freezed for tested . Immunofluorescence was used to detect FN and Pax7; The Real-time PCR was used to detect mRNA of FN, Sdc4, Fzd7, Wnt7a, c-Jun, Pax7; Western Blotting was used to detect the FN, sdc4, Fzd7, Wnt7a, c-Jun, p-c-Jun, Pax7 protein content. Results (1) Body weight, grip strength and skeletal muscle mass of mice: The body weight of group C and group R were significantly decreased after 9 weeks to compared with group A (P <0.05; P <0.05). The grip strength of group R and A was significantly increased in the ninth week to compared with group C, (P <0.01; P <0.05); The grip strength of group R and A in the ninth week were significantly higher than that in the first week (P <0.01; P <0.01)；  (2) FN in skeletal muscle extracellular matrix and it’s receptor Sdc4: The integrated optical density (IOD) of FN in group R was higher than that in group C and group A, but there was no significant difference among the three groups. The FN mRNA in group R was significantly increased to compared with group A (P<0.05). The FN protein content in group R was significantly increased to compared with group C and group A (P <0.01; P <0.01). There was no significant difference in the expression of Sdc4 mRNA in gastrocnemius muscle among the three groups. Compared with group C, the Sdc4 protein content was significantly down-regulated in both group R and group A (P<0.01; P<0.05). (3) Wnt7a/PCP signaling pathway: The Wnt7a mRNA and Wnt7a protein content in the gastrocnemius muscle of group R were significantly increased to compared with group C (P<0.05; P<0.01)). Compared with group A, Wnt7a protein content in group R was also significantly increased (P<0.05). There was no significant difference in Fzd7 mRNA in gastrocnemius muscle among the three groups; but the Fzd7 protein content in group R was significantly increased to compared with group A (P<0.05). The c-Jun mRNA in group R was significantly increased to compared with group C and A (P<0.05; P< 0.05). The content of c-Jun protein in group R was significantly increased to compared with group C and group A (P<0.01; P<0.01). The content of p-c-Jun protein in group R was significantly increased to compared with group C (P<0.05). (4) Pax7 : The number of Pax7 positive cells in group R was higher than that in group C and group A, but there was no significant difference among the three groups. Pax7 mRNA in group R and group A were significantly lower than that in group C (P<0.05; P<0.01), but Pax7 protein content in group R was significantly higher than group C and group A (P <0.05; P <0.05) Conclusions (1) Exercise can improve muscle strength of aging mice. (2) Resistance training can promote FN in skeletal muscle extracellular matrix and improve skeletal muscle extracellular matrix components of aging mice. (3) Resistance exercise training can promote Pax7 expression through the increase of FN and up-regulation of Wnt7a / PCP signaling pathway, thus make it possible for satellite cell proliferation.

scholarly journals Comparative RNA-sequencing profiled the differential gene expression of liver in response to acetyl-CoA carboxylase inhibitor GS-0976 in a mouse model of NASH

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e8115
Author(s):  
Ying Lu ◽  
Xiaolan Su ◽  
Manyu Zhao ◽  
Qianru Zhang ◽  
Chuang Liu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mouse Model ◽  
Rna Sequencing ◽  
Hepatic Steatosis ◽  
Differential Gene Expression ◽  
Receptor Interaction ◽  
Therapeutic Effects ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Differential Gene

Background Non-alcoholic steatohepatitis (NASH) is a progressive liver disease characterized by hepatic steatosis, lobular inflammation and fibrosis. Acetyl-CoA carboxylase (ACC) isoform 1 and 2 involved in de novo lipogenesis (DNL) and fatty acid oxidation have been identified as a therapeutic target in NASH. GS-0976, the inhibitor of ACC1 and ACC2, has achieved favorable therapeutic effects in clinical trials with NASH. The purpose of this study was to explore the transcriptional alterations regulated by GS-0976 in NASH. Methods C57BL/6 mice were fed on a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) or normal diet for 12 weeks. Mice were treated with or without GS-0976 (3 mg/kg per day) in the last 8 weeks. Oil Red O, Haematoxylin-eosin (H & E), and Sirius Red were used to evaluate hepatic steatosis, inflammation and fibrosis. The comparative RNA-sequencing was conducted to analyse the hepatic gene expression profiles in mice. Reverse transcription–polymerase chain reaction analysis was performed to validate the differential expression of representative genes. Results GS-0976 attenuated the steatosis, inflammation, and fibrosis of NASH in CDAHFD mouse model. High-throughput sequencing and differential gene expression analysis showed that there were 516 up-regulated genes and 525 down-regulated genes after GS-0976 treatment. Genes involved in the metabolic process, extracellular matrix formation, immune response, and angiogenesis were significantly enriched. The “Metabolic pathways” and “ECM-receptor interaction” pathways were the most significantly enriched KEGG pathways in the up-regulated and down-regulated differentially expressed genes (DEGs), respectively. Conclusions Transcriptome analysis showed that GS-0976 could regulate the expression of genes related to metabolism, inflammation and fibrosis in NASH. The global transcriptomic changes in gene expression promote the further understanding for the inhibition mechanisms of GS-0976 in NASH.

scholarly journals Pre-Operative Physical Function Predicts Post-Operative Skeletal Muscle Gene Expression in Old Mice

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. 830-830
Author(s):  
Samantha Asche-Godin ◽  
Lauren Harlow ◽  
Zachary Graham ◽  
Weihua Huang ◽  
Charles Mobbs ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Physical Function ◽  
Pathway Analysis ◽  
Sequencing Analysis ◽  
Muscle Gene Expression ◽  
Operative Outcomes ◽  
Whole Transcriptome Sequencing ◽  
Operative Performance ◽  
Muscle Gene

Abstract In older adults, pre-operative physical function predicts post-operative outcomes. The biological mechanisms underlying vulnerability to physical decline remain poorly understood. Using a mouse model of laparotomy, we sought to identify biological correlates of post-operative function. 24-month-old male C57BL/6N mice were categorized as high functioning (HF) or low functioning (LF) based on pre-operative performance on the accelerating rotarod. On post-operative days (POD) 2 and 4, LF mice had lower rotarod latency to fall times than HF mice did. Forelimb grip strength was reduced after laparotomy in both HF and LF groups on POD 1 and 3 and did not differ significantly between these groups. Whole transcriptome sequencing analysis (RNAseq) of soleus muscles collected on POD 5 showed 224 and 228 differentially expressed genes (DEGs) for HF and LF, respectively, compared to their respective controls. Only 21 DEGs were observed in both groups, including Pparα, Fst and Pla2g15. Such changes may be hallmarks of the post-surgical response in aging. Pathway analysis of DEGs using Ingenuity Pathway Analysis software (Qiagen) revealed one pathway common to HF and LF (osteoarthritis) whereas activation of GP6 signaling and apoptosis signaling was observed in HF and inhibition of PPARα/RXR activation and PPARα signaling was noted in LF. We conclude that pre-operative performance on the accelerating rotarod correlates with differences in skeletal muscle gene expression, which may contribute to the differences in functional outcomes post-operatively in HF and LF mice. Further studies are needed to delineate the roles of these signaling pathways in physical resilience to surgery.

Changes in uncoupling protein-2 and -3 expression in aging rat skeletal muscle, liver, and heart

2001 ◽  
Vol 280 (3) ◽  
pp. E413-E419 ◽  
Author(s):  
Rocco Barazzoni ◽  
K. Sreekumaran Nair
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Gastrocnemius Muscle ◽  
Uncoupling Protein ◽  
Mitochondrial Gene ◽  
Mtdna Copy Number ◽  
Transcript Levels ◽  
Atp Production ◽  
Age Related ◽  
Age Related Changes

Uncoupling protein (UCP)-2 and -3 mediate mitochondrial (mt) proton leak in vitro and are potential regulators of energy expenditure and ATP production. Aging is associated with alteration of tissue functions, suggesting impaired mtATP production. To determine whether age-related changes in UCP expression occur, we measured the transcript levels of UCP-2 and -3 in skeletal muscle, liver, and heart in 6- and 27-mo-old rats. UCP-2 transcripts were higher in old animals in the white (+100%) and red (+70%, both P < 0.04) gastrocnemius muscle and in the liver (+300%, P < 0.03), whereas they were comparable in the heart in both age groups. UCP-2 transcript levels correlated positively with mitochondrial-encoded cytochrome c oxidase transcripts normalized for mtDNA ( P < 0.01) and negatively with mtDNA copy number ( P < 0.001). UCP-3 transcripts were lower in the less oxidative white (−50%, P < 0.04) and unchanged in the more oxidative red (−15%, P = 0.41) gastrocnemius muscle in old animals. Similar changes at protein level were confirmed by UCP-2 protein in aging liver (+300%, P < 0.01) and UCP-2 (+85%, P < 0.05) and UCP-3 (−30%, P = 0.4) protein in aging mixed gastrocnemius muscle. Aging is thus associated with tissue-specific changes of UCP-2 and -3 gene expression. Increased UCP-2 expression may limit ATP production and is related to mitochondrial gene expression in aging muscles and liver. Different age-related changes may reflect differential regulation of UCP-2 and -3 in skeletal muscle. The current data suggest a potential role of uncoupling proteins to alter energy production in aging tissues.

scholarly journals Transcriptome-wide RNA sequencing analysis of rat skeletal muscle feed arteries. II. Impact of exercise training in obesity

2014 ◽  
Vol 116 (8) ◽  
pp. 1033-1047 ◽  
Author(s):  
Jaume Padilla ◽  
Nathan T. Jenkins ◽  
Pamela K. Thorne ◽  
Jeffrey S. Martin ◽  
R. Scott Rector ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Exercise Training ◽  
Training Program ◽  
Rna Sequencing ◽  
Gastrocnemius Muscle ◽  
Aortic Endothelial Cells ◽  
Feed Arteries ◽  
Skeletal Muscle Feed Arteries ◽  
Aortic Endothelial

We employed next-generation RNA sequencing (RNA-Seq) technology to determine the extent to which exercise training alters global gene expression in skeletal muscle feed arteries and aortic endothelial cells of obese Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Transcriptional profiles of the soleus and gastrocnemius muscle feed arteries (SFA and GFA, respectively) and aortic endothelial cell-enriched samples from rats that underwent an endurance exercise training program (EndEx; n = 12) or a interval sprint training program (IST; n = 12) or remained sedentary (Sed; n = 12) were examined. In response to EndEx, there were 39 upregulated (e.g., MANF) and 20 downregulated (e.g., ALOX15) genes in SFA and 1 upregulated (i.e., Wisp2) and 1 downregulated (i.e., Crem) gene in GFA [false discovery rate (FDR) < 10%]. In response to IST, there were 305 upregulated (e.g., MANF, HSPA12B) and 324 downregulated genes in SFA and 101 upregulated and 66 downregulated genes in GFA, with an overlap of 32 genes between arteries. Furthermore, in aortic endothelial cells, there were 183 upregulated (e.g., eNOS, SOD-3) and 141 downregulated (e.g., ATF3, Clec1b, npy, leptin) genes with EndEx and 71 upregulated and 69 downregulated genes with IST, with an overlap of 35 between exercise programs. Expression of only two genes (Tubb2b and Slc9a3r2) was altered (i.e., increased) by exercise in all three arteries. The finding that both EndEx and IST produced greater transcriptional changes in the SFA compared with the GFA is intriguing when considering the fact that treadmill bouts of exercise are associated with greater relative increases in blood flow to the gastrocnemius muscle compared with the soleus muscle.

In vivo regulation of SREBP-1c in skeletal muscle: effects of nutritional status, glucose, insulin, and leptin

2004 ◽  
Vol 287 (1) ◽  
pp. R218-R227 ◽  
Author(s):  
S. Renee Commerford ◽  
Li Peng ◽  
John J. Dubé ◽  
Robert M. O'Doherty
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Nutritional Status ◽  
Fatty Acid Synthase ◽  
Gastrocnemius Muscle ◽  
Uncoupling Protein ◽  
Regulatory Element ◽  
Mrna Level ◽  
Mrna Levels ◽  
Fas Mrna

Sterol regulatory element binding protein-1c (SREBP-1c), a transcription factor that is important for mediating insulin effects on metabolic gene expression in liver during the fasted-to-fed transition, is also expressed in skeletal muscle. However, the regulation and role of SREBP-1c in skeletal muscle are poorly understood. The present study compared the effects of nutritional status, physiological hyperinsulinemic clamps, and adenovirus-mediated hyperleptinemia (HLEP) in rats on expression of SREBP-1c and other metabolic genes in skeletal muscle. Three- and 6-h refeeding of 18-h-fasted animals increased levels of SREBP-1c mRNA and the SREBP-1 protein (full length and mature) in gastrocnemius muscle ( P < 0.05). Fatty acid synthase (FAS) and hexokinase II (HKII) mRNA levels were also increased by refeeding, and uncoupling protein 3 (UCP3) mRNA level was decreased (all P < 0.05). Surprisingly, 3-h hyperinsulinemic clamps did not increase gastrocnemius muscle SREBP-1c and FAS mRNA levels or SREBP-1 protein levels but did increase HKII mRNA levels and decrease UCP3 mRNA levels ( P < 0.05). HLEP reduced refeeding-induced increases of SREBP-1c and FAS mRNA levels but did not reduce the level of SREBP-1 protein. We conclude that 1) skeletal muscle SREBP-1c gene expression is regulated by nutritional status in a fashion similar to that observed in liver and adipose tissue, 2) physiological hyperinsulinemia is not sufficient to imitate the effects of refeeding on SREBP-1c gene expression, and 3) leptin suppresses refeeding effects on SREBP-1c mRNA levels.

scholarly journals Single-cell Bottlenecks and Dead-ends During Glioma Premalignancy

2020 ◽  
Author(s):  
Matthew D. Sutcliffe ◽  
Rui P. Galvao ◽  
Lixin Wang ◽  
Jungeun Kim ◽  
Shambhavi Singh ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mouse Model ◽  
Rna Sequencing ◽  
Replication Stress ◽  
Fluctuation Analysis ◽  
Global Changes ◽  
Differential Analysis ◽  
Cell Of Origin ◽  
Cell State ◽  
Genetically Engineered Mouse Model

AbstractCancer evolves from premalignant clones that accumulate mutations and adopt unusual cell states to achieve transformation. Tracking a cancer cell-of-origin through the cell-state alterations of premalignancy could provide clues for early-detection and cancer-prevention strategies. Previously we pinpointed the oligodendrocyte precursor cell (OPC) as a cell-of-origin for glioma. However, the early adaptations and cell-state changes of mutant OPCs during premalignancy are unknown. Using a genetically engineered mouse model (GEMM) of inducible Nf1–Trp53 loss in OPCs, we acutely isolated labeled mutant OPCs by laser-capture microdissection and determined gene-expression changes in two ways: global changes in gene expression were measured by differential analysis of wild-type and mutant OPCs after bulk RNA sequencing; cell-to-cell state variations were identified by a fluctuation analysis, called stochastic profiling, which uses RNA-sequencing measurements from random pools of 10 mutant cells. We chose two time points for the analysis. At 12 days after Nf1–Trp53 deletion, while bulk differences were mostly limited to increases in mitotic hallmarks and decreases in ribosome biosynthesis, stochastic profiling of mutant OPCs revealed a spectrum of stem-progenitor, proneural, and mesenchymal states as potential starting points for gliomagenesis. At 90 days after Nf1–Trp53 deletion, while bulk sequencing detected very few differentially expressed transcripts, stochastic profiling revealed multiple cell states that are absent from glial tumors, suggesting that they marked dead-ends for gliomagenesis. In parallel, we identified cells without dead-end markers but abundantly expressing key effectors of nonsense-mediated decay and homology-dependent DNA repair. This suggests that resolution of replication stress may pose a considerable bottleneck for glioma initiation in premalignant mutant OPCs.Statement of significanceIn situ heterogeneity profiling of cell states in a mouse model of glioma uncovers regulatory confusion in a glioma cell-of-origin and defines a state of replication stress that precedes tumor initiation.

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