scholarly journals Histochemical Characterisation and Gene Expression Analysis of Skeletal Muscles from Maremmana and Aubrac Steers Reared on Grazing and Feedlot Systems

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 656
Author(s):  
Giulia Foggi ◽  
Francesca Ciucci ◽  
Maria Conte ◽  
Laura Casarosa ◽  
Andrea Serra ◽  
...  
Keyword(s):  
Gene Expression ◽  
Physical Activity ◽  
Muscle Fibre ◽  
Skeletal Muscles ◽  
Gene Expression Analysis ◽  
Type I ◽  
Triceps Brachii ◽  
Gene Expressions ◽  
Muscle Properties ◽  
Voluntary Physical Activity

This study aimed to characterise the fibre composition of Triceps brachii (TB) and Semimembranosus (SM) muscles from 20 Maremmana (MA) and 20 Aubrac (AU) steers, and the effect of grazing activity in comparison with feedlot system. The histochemical method was performed with the m-ATPase method with an acid pre-incubation, thus allowing to distinguish type I, IIA, and IIB fibres. Additionally, on total RNA extracted from SM muscle, the expressions of atp1a1, mt-atp6, and capn1 genes were evaluated, in order to find potential associations with muscle fibre histochemical characteristics. In SM muscle, the MA steers had the greater frequency of oxidative fibres (type I and IIA) and the higher atp1a1 expression, in comparison to AU steers. Conversely, AU steers had a greater frequency of type IIB fibres, and the higher capn1 expression. A similar histochemical pattern was observed in TB muscle. The grazing activity was probably insufficient to determine differences both for fibre proportion and size, and gene expressions, except for mt-atp6 expression that was surprisingly highest in feedlot MA in comparison to other steers. These findings further the knowledge of muscle properties belonging to these breeds, and the effect of voluntary physical activity since few studies were available in this regard.

scholarly journals Analysis of Activity-Dependent Energy Metabolism in Mice Reveals Regulation of Mitochondrial Fission and Fusion mRNA by Voluntary Physical Exercise in Subcutaneous Fat from Male Marathon Mice (DUhTP)

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2697
Author(s):  
Julia Brenmoehl ◽  
Daniela Ohde ◽  
Christina Walz ◽  
Martina Langhammer ◽  
Julia Schultz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Physical Activity ◽  
Energy Metabolism ◽  
Fat Mass ◽  
Mitochondrial Fission ◽  
Subcutaneous Fat ◽  
Voluntary Exercise ◽  
Heart Weight ◽  
Mitochondrial Energy Metabolism ◽  
Voluntary Physical Activity

Physical inactivity is considered as one of the main causes of obesity in modern civilizations, and it has been demonstrated that resistance training programs can be used to reduce fat mass. The effects of voluntary exercise on energy metabolism are less clear in adipose tissue. Therefore, the effects of three different voluntary exercise programs on the control of energy metabolism in subcutaneous fat were tested in two different mouse lines. In a cross-over study design, male mice were kept for three or six weeks in the presence or absence of running wheels. For the experiment, mice with increased running capacity (DUhTP) were used and compared to controls (DUC). Body and organ weight, feed intake, and voluntary running wheel activity were recorded. In subcutaneous fat, gene expression of browning markers and mitochondrial energy metabolism were analyzed. Exercise increased heart weight in control mice (p < 0.05) but significantly decreased subcutaneous, epididymal, perinephric, and brown fat mass in both genetic groups (p < 0.05). Gene expression analysis revealed higher expression of browning markers and individual complex subunits present in the electron transport chain in subcutaneous fat of DUhTP mice compared to controls (DUC; p < 0.01), independent of physical activity. While in control mice, voluntary exercise had no effect on markers of mitochondrial fission or fusion, in DUhTP mice, reduced mitochondrial DNA, transcription factor Nrf1, fission- (Dnm1), and fusion-relevant transcripts (Mfn1 and 2) were observed in response to voluntary physical activity (p < 0.05). Our findings indicate that the superior running abilities in DUhTP mice, on one hand, are connected to elevated expression of genetic markers for browning and oxidative phosphorylation in subcutaneous fat. In subcutaneous fat from DUhTP but not in unselected control mice, we further demonstrate reduced expression of genes for mitochondrial fission and fusion in response to voluntary physical activity.

Inverse alterations of BCKA dehydrogenase activity in cardiac and skeletal muscles of diabetic rats

1999 ◽  
Vol 277 (4) ◽  
pp. E685-E692 ◽  
Author(s):  
Yolanda B. Lombardo ◽  
Cynthia Serdikoff ◽  
Manikkavasagar Thamotharan ◽  
Harbhajan S. Paul ◽  
Siamak A. Adibi
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Protein Expression ◽  
Cardiac Muscle ◽  
Dehydrogenase Activity ◽  
Skeletal Muscles ◽  
Diabetic Rats ◽  
Gene Expressions ◽  
The Difference ◽  
Activity State

Rat cardiac and skeletal muscles, which have been used as model tissues for studies of regulation of branched-chain α-keto acid (BCKA) oxidation, vary greatly in the activity state of their BCKA dehydrogenase. In the present experiment, we have investigated whether they also vary in response of their BCKA dehydrogenase to a metabolic alteration such as diabetes and, if so, to investigate the mechanism that underlies the difference. Diabetes was produced by depriving streptozotocin-treated rats of insulin administration for 96 h. The investigation of BCKA dehydrogenase in the skeletal muscle (gastrocnemius) showed that diabetes 1) increased its activity, 2) increased the protein and gene expressions of all of its subunits (E1α, E1β, E2), 3) increased its activity state, 4) decreased the rate of its inactivation, and 5) decreased the protein expression of its associated kinase (BCKAD kinase) without affecting its gene expression. In sharp contrast, the investigation of BCKA dehydrogenase in the cardiac muscle showed that diabetes 1) decreased its activity, 2) had no effect on either protein or gene expression of any of its subunits, 3) decreased its activity state, 4) increased its rate of inactivation, and 5) increased both the protein and gene expressions of its associated kinase. In conclusion, our data suggest that, in diabetes, the protein expression of BCKAD kinase is downregulated posttranscriptionally in the skeletal muscle, whereas it is upregulated pretranslationally in the cardiac muscle, causing inverse alterations of BCKA dehydrogenase activity in these muscles.

scholarly journals An Integrated Preprocessing Approach for Exploring Single-Cell Gene Expression in Rare Cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Junyi Shang ◽  
David Welch ◽  
Manuela Buonanno ◽  
Brian Ponnaiya ◽  
Guy Garty ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Bystander Effect ◽  
Kinase Inhibitor ◽  
Digital Pcr ◽  
Lung Fibroblasts ◽  
Gene Expressions ◽  
Rare Cells

AbstractExploring the variability in gene expressions of rare cells at the single-cell level is critical for understanding mechanisms of differentiation in tissue function and development as well as for disease diagnostics and cancer treatment. Such studies, however, have been hindered by major difficulties in tracking the identity of individual cells. We present an approach that combines single-cell picking, lysing, reverse transcription and digital polymerase chain reaction to enable the isolation, tracking and gene expression analysis of rare cells. The approach utilizes a photocleavage bead-based microfluidic device to synthesize and deliver stable cDNA for downstream gene expression analysis, thereby allowing chip-based integration of multiple reactions and facilitating the minimization of sample loss or contamination. The utility of the approach was demonstrated with QuantStudio digital PCR by analyzing the radiation and bystander effect on individual IMR90 human lung fibroblasts. Expression levels of the Cyclin-dependent kinase inhibitor 1a (CDKN1A), Growth/differentiation factor 15 (GDF15), and Prostaglandin-endoperoxide synthase 2 (PTGS2) genes, previously shown to have different responses to direct and bystander irradiation, were measured across individual control, microbeam-irradiated or bystander IMR90 cells. In addition to the confirmation of accurate tracking of cell treatments through the system and efficient analysis of single-cell responses, the results enable comparison of activation levels of different genes and provide insight into signaling pathways within individual cells.

scholarly journals Ractopamine-induced fiber type-specific gene expression in porcine skeletal muscles is independent of growth

2020 ◽  
Vol 98 (11) ◽  
Author(s):  
Andrea M Gunawan ◽  
Con-Ning Yen ◽  
Brian T Richert ◽  
Allan P Schinckel ◽  
Alan L Grant ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscles ◽  
Muscle Hypertrophy ◽  
Citrate Synthase ◽  
Fiber Type ◽  
Muscle Growth ◽  
Specific Gene ◽  
Type I ◽  
Fiber Types ◽  
Adequate Diet

Abstract Feeding ractopamine (RAC), a β-adrenergic agonist (BAA), to pigs increases type IIB muscle fiber type-specific protein and mRNA expression. However, increases in the abundance of these fast-twitch fiber types occur with other forms of muscle hypertrophy and thus BAA-induced changes in myosin heavy chain (MyHC) composition may simply be associated with increased muscle growth known to occur in response to BAA feeding. The objective of this study was to determine whether RAC feeding could change the MyHC gene expression in the absence of maximal muscle growth. Pigs were fed either an adequate diet that supported maximal muscle hypertrophy or a low nutrient diet that limited muscle growth. RAC was included in diets at 0 or 20 mg/kg for 1, 2, or 4 wk. Backfat depth was less (P &lt; 0.05) in pigs fed the low nutrient diet compared with the adequate diet but was not affected by RAC. Loin eye area was greater (P &lt; 0.05) in pigs fed an adequate diet plus RAC at 1 wk but did not differ among remaining pigs. At 2 and 4 wk, however, pigs fed the adequate diet had greater loin eye areas (P &lt; 0.05) than pigs fed the low nutrient diet regardless of RAC feeding. Gene expression of the MyHC isoforms, I, IIA, IIX, and IIB, as well as glycogen synthase, citrate synthase, β 1-adrenergic receptor (AR), and β 2-AR were determined in longissimus dorsi (LD) and red (RST) and white (WST) portions of the semitendinosus muscles. MyHC type I gene expression was not altered by RAC or diet. Feeding RAC decreased (P &lt; 0.01) MyHC type IIA gene expression in all muscles, but to a greater extent in WST and LD. MyHC type IIX gene expression was lower (P &lt; 0.05) in WST and LD muscles in response to RAC but was not altered in RST muscles. RAC increased (P &lt; 0.05) MyHC type IIB gene expression in all muscles, but to a greater extent in RST. β 1-AR gene expression was unaffected by RAC or diet, whereas the expression of the β 2-AR gene was decreased (P &lt; 0.001) by RAC. No significant RAC * diet interactions were observed in gene expression in this study, indicating that RAC altered MyHC and β 2-AR gene expression in porcine skeletal muscles independent of growth.

Effects of Controlled Released BMP-7 on Markers of Inflammation and Degradation During the Cultivation of Human Osteoarthritic Chondrocytes

2010 ◽  
Vol 26 (4) ◽  
pp. 419-433 ◽  
Author(s):  
Karsten Gavenis ◽  
Thomas Pufe ◽  
Lars Ove Brandenburg ◽  
Katharina Schiffl ◽  
Bernhard Schmidt-Rohlfing
Keyword(s):  
Gene Expression ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Type I ◽  
Matrix Synthesis ◽  
Gene Expressions ◽  
Markers Of Inflammation ◽  
Osteoarthritic Chondrocytes

The aim of the present study is to investigate the effects of BMP-7 released from polylactide microspheres on the appearance of various catabolic and inflammatory cytokines secreted by osteoarthritic chondrocytes cultivated in a collagen gel. Articular chondrocytes of 15 patients suffering from osteoarthritis are transferred to a collagen type-I gel. Additionally, BMP-7 encapsulated into polylactide microspheres (50 ng BMP-7/mL gel) is added. After 14 days, gene expression and protein appearance of various genes involved in matrix turnover and inflammation are investigated by immunohistochemical staining and RT-PCR and compared to untreated controls. TNF-α, MMP-13, IL-6, IL-1β, and VEGF gene expressions are decreased in the treatment group. In contrast, BMP-7-induced matrix synthesis is not affected, leaving collagen type-II (Col-II) gene expression to be elevated, while collagen type-I (Col-I) is decreased. In summary, controlled release of low concentrated BMP-7 from polylactide microspheres leads to a decrease in gene expression of the investigated inflammation and matrix degradation markers whereas matrix synthesis is induced.

scholarly journals Voluntary exercise and testosterone therapy caused increase in percentage of Myh6 and expression of oxidative stress marker Cybb in left ventricles of rats

2016 ◽  
Vol 63 (1) ◽  
pp. 12-15 ◽  
Author(s):  
M. Radik ◽  
G. Doka ◽  
E. Malikova ◽  
P. Krenek ◽  
J. Klimas
Keyword(s):  
Oxidative Stress ◽  
Physical Activity ◽  
Polymerase Chain Reaction Method ◽  
Voluntary Exercise ◽  
Oxidative Stress Marker ◽  
Gene Expressions ◽  
Myosin Heavy Chain Isoform ◽  
Expression Of Genes ◽  
Voluntary Physical Activity ◽  
Male Wistar Rats

Abstract Aim: The aim of this study is to identify a possible damage to heart ventricles caused by supraphysiological doses of testosterone, voluntary physical activity or their combination. Methods: In the 8-week long experiment, 10-12 weeks old male Wistar rats were administered testosterone depot in dose of 100 mg/kg (TES, n = 15) or vehiculum (CON, n = 12) once a week subcutaneously. Next groups injected with testosterone (SPOTES, n = 12) or vehiculum (SPO, n = 12) were running in exercise wheels ad libitum. Gene expressions in left and right ventricles of the heart were measured by quantitative reverse transcription polymerase chain reaction method. Results:ln left ventricles of the testosterone groups, we observed a mild but significant increase in the percentage of Myh6 myosin heavy chain isoform and higher expression of NADPH oxidase subunit Cybb (*p < 0.05). Conclusions:Testosterone affected the expression of genes related to contractile apparatus and oxidative stress in the left ventricle but not in right ventricle of the heart of rats. The observed level of physical activity did not have a compelling effect on the expression of measured genes.

scholarly journals Anandamide Influences Interleukin-1β Synthesis and IL-1 System Gene Expressions in the Ovine Hypothalamus during Endo-Toxin-Induced Inflammation

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 484
Author(s):  
Monika Tomczyk ◽  
Dorota Tomaszewska-Zaremba ◽  
Joanna Bochenek ◽  
Anna Herman ◽  
Andrzej P. Herman
Keyword(s):  
Gene Expression ◽  
Central Nervous System ◽  
Nervous System ◽  
Receptor Antagonist ◽  
Inflammatory Process ◽  
Type I ◽  
Central Control ◽  
Gene Expressions ◽  
Cytokine Synthesis ◽  
Inflammatory Effect

This study evaluated the effect of anandamide (AEA) on interleukin (IL)-1β synthesis and gene expression of IL-1β, its type I (IL-1R1) and II (IL-1R2) receptors, and IL-1 receptor antagonist (IL-1RN) in the hypothalamic structures, involved in the central control of reproduction, during inflammation. Animals were intravenously (i.v.) injected with bacterial endotoxin-lipopolysaccharide (LPS) (400 ng/kg) or saline, and two hours after LPS administration., a third group received i.v. injection of AEA (10 μg/kg). Ewes were euthanized one hour later. AEA injection (p < 0.05) suppressed LPS-induced expression of IL-1β protein in the hypothalamus. The gene expression of IL-1β, IL-1RN, and IL-1R2 in the hypothalamic structures was higher (p < 0.05) in animals treated with both LPS and AEA in comparison to other experimental groups. AEA administration did not influence LPS-stimulated IL-1R1 gene expression. Our study shows that AEA suppressed IL-1β synthesis in the hypothalamus, likely affecting posttranscriptional levels of this cytokine synthesis. However, anti-inflammatory effect of AEA might also result from its stimulating action on IL-1RN and IL-1R2 gene expression. These results indicate the potential of endocannabinoids and/or their metabolites in the inhibition of inflammatory process at the level of central nervous system, and therefore their usefulness in the therapy of inflammation-induced neuroendocrine disorders.

Annatto (Bixa orellana) δ-TCT supplementation protected against embryonic DNA damages through alterations in PI3K/ Akt-Cyclin D1 pathway

2018 ◽  
Vol 88 (1-2) ◽  
pp. 16-26
Author(s):  
Siti Syairah Mohd Mutalip ◽  
Mohd Hamim Rajikin ◽  
Sharaniza Ab Rahim ◽  
Norashikin Mohamed Noor Khan
Keyword(s):  
Gene Expression ◽  
Cyclin D1 ◽  
Gene Expression Analysis ◽  
Protective Action ◽  
Alpha Tocopherol ◽  
Bixa Orellana ◽  
Inhibitory Effects ◽  
Gene Expressions ◽  
Concurrent Treatment ◽  
Dna Damages

Abstract. Protective action by annatto-derived delta-tocotrienol (δ-TCT) and soy-derived alpha-tocopherol (α-TOC) through the regulation of PI3K/Akt-Cyclin D1 pathway against the nicotine-induced DNA damages is the focus of the present study. Nicotine, which has been widely reported to have numerous adverse effects on the reproductive system, was used as reproductive toxicant. 48 female balb/c mice (6–8 weeks) (23–25 g) were randomly divided into 8 groups (G1-G8; n = 6) and treated with either nicotine or/and annatto δ-TCT/soy α-TOC for 7 consecutive days. On Day 8, the females were superovulated and mated before euthanized for embryo collection (46 hours post-coitum). Fifty 2-cell embryos from each group were used in gene expression analysis using Affymetrix QuantiGene Plex2.0 assay. Findings indicated that nicotine (G2) significantly decreased (p < 0.05) the number of produced 2-cell embryos compared to control (G1). Intervention with mixed annatto δ-TCT (G3) and pure annatto δ-TCT (G4) significantly increased the number of produced 2-cell embryos by 127 % and 79 % respectively compared to G2, but these were lower than G1. Concurrent treatment with soy α-TOC (G5) decreased embryo production by 7 %. Supplementations with δ-TCT and α-TOC alone (G6-G8) significantly increased (p < 0.05) the number of produced 2-cell embryos by 50 %, 36 % and 41 % respectively, compared to control (G1). These results were found to be associated with the alterations in the PI3K/Akt-Cyclin D1 gene expressions, indicating the inhibitory effects of annatto δ-TCT and soy α-TOC against the nicotinic embryonic damages. To our knowledge, this is the first attempt on studying the benefits of annatto δ-TCT on murine preimplantation 2-cell embryos.

scholarly journals Enhanced Growth of Lapine Anterior Cruciate Ligament-Derived Fibroblasts on Scaffolds Embroidered from Poly(l-lactide-co-ε-caprolactone) and Polylactic Acid Threads Functionalized by Fluorination and Hexamethylene Diisocyanate Cross-Linked Collagen Foams

2020 ◽  
Vol 21 (3) ◽  
pp. 1132 ◽  
Author(s):  
Clemens Gögele ◽  
Judith Hahn ◽  
Cindy Elschner ◽  
Annette Breier ◽  
Michaela Schröpfer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tissue Engineering ◽  
Polylactic Acid ◽  
Cruciate Ligament ◽  
Donor Site ◽  
Hexamethylene Diisocyanate ◽  
Type I ◽  
Gene Expressions ◽  
Extracellular Matrix Components ◽  
Anterior Cruciate

Reconstruction of ruptured anterior cruciate ligaments (ACLs) is limited by the availability and donor site morbidity of autografts. Hence, a tissue engineered graft could present an alternative in the future. This study was undertaken to determine the performance of lapine (L) ACL-derived fibroblasts on embroidered poly(l-lactide-co-ε-caprolactone) (P(LA-CL)) and polylactic acid (PLA) scaffolds in regard to a tissue engineering approach for ACL reconstruction. Surface modifications of P(LA-CL)/PLA by gas-phase fluorination and cross-linking of a collagen foam using either ethylcarbodiimide (EDC) or hexamethylene diisocyanate (HMDI) were tested regarding their influence on cell adhesion, growth and gene expression. The experiments were performed using embroidered P(LA-CL)/PLA scaffolds that were seeded dynamically or statically with LACL-derived fibroblasts. Scaffold cytocompatibility, cell survival, numbers, metabolic activity, ultrastructure and sulfated glycosaminoglycan (sGAG) synthesis were evaluated. Quantitative real-time polymerase chain reaction (QPCR) revealed gene expression of collagen type I (COL1A1), decorin (DCN), tenascin C (TNC), Mohawk (MKX) and tenomodulin (TNMD). All tested scaffolds were highly cytocompatible. A significantly higher cellularity and larger scaffold surface areas colonized by cells were detected in HMDI cross-linked and fluorinated scaffolds compared to those cross-linked with EDC or without any functionalization. By contrast, sGAG synthesis was higher in controls. Despite the fact that the significance level was not reached, gene expressions of ligament extracellular matrix components and differentiation markers were generally higher in fluorinated scaffolds with cross-linked collagen foams. LACL-derived fibroblasts maintained their differentiated phenotype on fluorinated scaffolds supplemented with a HMDI cross-linked collagen foam, making them a promising tool for ACL tissue engineering.

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