ABSTRACT
Proteomics plays a key and insightful role in meat research in the post-genomic era. This study aimed to unveil using a shotgun proteomics approach the temporal dynamic changes in early post-mortem proteome of goat Semitendinosus muscle. Therefore, the evolution and comparison of the muscle proteome over three post-mortem times (1, 8, and 24 h) was assessed. The temporal proteomics profiling quantified 748 proteins, from which 174 were differentially abundant (DAPs): n = 55 between 1 h versus 8 h, n = 52 between 8 h versus 24 h, and n = 154 between 1 h versus 24 h. The DAPs belong to myriad interconnected pathways. Binding, transport and calcium homeostasis, as well as muscle contraction and structure, exhibited an equivalent contribution during post-mortem, demonstrating their central role. Catalytic, metabolism and ATP metabolic process, and proteolysis were active pathways from the first hours of animal bleeding. Conversely, oxidative stress, response to hypoxia and cell redox homeostasis along chaperones and heat shock proteins accounted for the large proportion of the biochemical processes, more importantly after 8 h post-mortem. Overall, the conversion of muscle into meat is largely orchestrated by energy production as well as mitochondrial metabolism and homeostasis through calcium and permeability transition regulation. The study further evidenced the role of ribosomal proteins in goat post-mortem muscle, signifying that several proteins experiencing changes during storage, also undergo splicing modifications, which is for instance a mechanism known for mitochondrial proteins. Overall, temporal proteomics profiling of early post-mortem muscle proteome offers an unparalleled view of the sophisticated post-mortem biochemical and proteolytic events associated with goat meat quality determination.
Subject(s)
Hamstring Muscles , Muscle Proteins , Animals , Muscle Proteins/metabolism , Proteome , Goats/metabolism , Hamstring Muscles/metabolism , Proteomics , Calcium/metabolism , Muscles/metabolism , Meat/analysis , Muscle, Skeletal/metabolism , Postmortem ChangesABSTRACT
Intramuscular fat content is an important factor that determines meat quality in pigs. In recent years, epigenetic regulation has increasingly studied the physiological model of intramuscular fat. Although long noncoding RNAs (lncRNAs) play essential roles in various biological processes, their role in intramuscular fat deposition in pigs remains largely unknown. In this study, intramuscular preadipocytes in the longissimus dorsi and semitendinosus of Large White pigs were isolated and induced into adipogenic differentiation in vitro. High-throughput RNA-seq was carried out to estimate the expression of lncRNAs at 0, 2 and 8 days post-differentiation. At this stage, 2135 lncRNAs were identified. KEGG analysis showed that the differentially expressed lncRNAs were common in pathways involved with adipogenesis and lipid metabolism. lnc_000368 was found to gradually increase during the adipogenic process. Reverse-transcription quantitative polymerase chain reaction and a western blot revealed that the knockdown of lnc_000368 significantly repressed the expression of adipogenic genes and lipolytic genes. As a result, lipid accumulation in porcine intramuscular adipocytes was impaired by the silencing of lnc_000368. Overall, our study identified a genome-wide lncRNA profile related to porcine intramuscular fat deposition, and the results suggest that lnc_000368 is a potential target gene that might be targeted in pig breeding in the future.
Subject(s)
Hamstring Muscles , RNA, Long Noncoding , Swine/genetics , Animals , Adipogenesis/genetics , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Hamstring Muscles/metabolism , Epigenesis, Genetic , Adipocytes/metabolismABSTRACT
14C released from nuclear facilities is transferred to cattle through their consumption of 14C contaminated grasses. To estimate the concentrations of 14C in their meat, we conducted two sets of experiments. In the first experiment, 230 mg of 13C per day was administered to cattle aged 10 months for 28 days in the form of 13C-labeled grass. The 13C concentration in the semitendinosus muscle decreased exponentially after reaching its peak value. The mean half-life was 76 ± 13 days. In the second experiment, 550 mg of 13C per day was administered to 24-month-old cattle. The change in the semitendinosus muscle was smaller than that recorded in the first experiment, even though the amount of 13C administered per body weight was slightly higher than that in the first experiment. Consequently, the half-life was not determined. Therefore, further studies are required to clarify the metabolism of carbon in 2-year-old cattle.
Subject(s)
Dactylis , Hamstring Muscles , Animals , Carbon , Carbon Isotopes , Cattle , Dactylis/metabolism , Hamstring Muscles/metabolismABSTRACT
This paper aimed to evaluate the effects of ultrasound-assisted L-histidine marination (UMH) on meat quality and actomyosin properties of beef M. semitendinosus. Our results found that UMH treatment effectively avoided excessive liquid withdrawal, and disrupted myofibril integrity by modifying the water distribution and weakening connection of actin-myosin with increased muscle pH. The ultrasound-treated sample provided more opportunity for the filtration of L-histidine to intervene the isoelectric point and conformation of muscle protein. The activated caspase-3 and changes of ATPase activity in UMH-treated meat accelerated the postmortem ageing, and L-histidine might competitively inhibit the actin-myosin binding by the imidazole group. UMH decreased the surface hydrophobicity by shielding hydrophobic area and unfolding the actomyosin structure. In addition, the increased actomyosin solubility with smaller particle size enhanced the SH content for better cross-linking of myosin tail, and formation of heat-set gelling protein structure. Therefore, UMH treatment manifested the potential to improve beef quality.
Subject(s)
Actomyosin , Hamstring Muscles , Actins , Actomyosin/chemistry , Animals , Cattle , Hamstring Muscles/metabolism , Histidine/chemistry , Meat/analysis , Myosins/chemistryABSTRACT
This study was conceived to evaluate the variations between backfat (BF) and Semimembranosus (SM) muscle fatty acid (FA) composition in 789 heavy pigs and to estimate the genetic and phenotypic correlations and the heritability values of these variations. Most FAs showed a common genetic basis controlling their proportion in SM muscle and BF, while the n-6/n-3 ratio, α-linolenic and erucic acids displayed a genetic control more oriented towards tissue-specific molecular pathways. All variations between the FA composition of BF and SM muscle showed low-to-medium heritability values, suggesting that there are also genetic mechanisms capable of differentiating the deposition of FAs in BF from those in SM muscle. This result implies that a better knowledge of the genes differentiating the FA composition of BF and SM muscle could provide new tools allowing to select, in a partially independent manner, the FA composition of muscle and subcutaneous fat.
Subject(s)
Fatty Acids , Hamstring Muscles , Adipose Tissue/metabolism , Animals , Body Composition/genetics , Fatty Acids/metabolism , Gene Expression Regulation , Hamstring Muscles/metabolism , Meat/analysis , Muscle, Skeletal/metabolism , Subcutaneous Fat/metabolism , Swine/geneticsABSTRACT
Glycolytic potential (GP) is one of the postmortem traits used to predict the quality of the final meat products. Despite that, the knowledge of the molecular and metabolic pathways controlling this trait is still not complete. To add some information on this field we used two pools of Italian Large White heavy pigs divergent for GP to investigate through a microarray approach the differences of gene expressions between the two pools. On the whole, 32 genes were differentially expressed, and among them 31 were overexpressed in low GP pool. These genes were involved in mitochondrial functions and adenosine triphosphate (ATP) biosynthetic processes, in calcium homeostasis, and in lipid metabolism, with Peroxisome proliferator-activated receptor (PPAR) signaling being a possible master regulator of the molecular differences observed between the two pools. The different GP levels between the two pools could have determined in low GP muscles a more rapid occurrence of the molecular cascade related to the events triggering cell death.
Subject(s)
Hamstring Muscles , Meat , Animals , Glycolysis/genetics , Hamstring Muscles/metabolism , Lipid Metabolism/genetics , Meat/analysis , Muscle, Skeletal/metabolism , Swine/geneticsABSTRACT
Towards a better understanding of the formation mechanism of salt on intramuscular triglyceride (TG) hydrolysis occurring in biceps femoris (BF) muscles during dry-salting process, the changes of TG hydrolysis, TG hydrolysis activity and phosphorylation of adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL) as well as their regulatory proteins (Perilipin1, ABHD5, G0S2) with different salt content (0%, 1%, 3%, 5%) and salting time (the first and third day) were analyzed. The results showed that dry-salting significantly increased the TG hydrolase activity and hydrolysis extent with salting process proceed (P < 0.05), especially upon the treatment with 3% amount of salt. The SDS-PAGE and Western-blot results further demonstrated that the promotion of salt on TG hydrolysis in intramuscular adipocytes was mainly attributed to the activation of protein kinase activity and protein phosphorylation process. Accordingly, the ATGL and HSL were activated, and meanwhile, the TG hydrolysis pivotal switch perilipin1 was also turned on by phosphorylation modification.
Subject(s)
Hamstring Muscles , Sterol Esterase , Animals , Hamstring Muscles/metabolism , Hydrolysis , Lipase/genetics , Lipase/metabolism , Lipids , Lipolysis , Phosphorylation , Sterol Esterase/genetics , Sterol Esterase/metabolism , Swine , TriglyceridesABSTRACT
The myotendinous junction (MTJ) is a specialized interface for transmitting high forces between the muscle and tendon and yet the MTJ is a common site of strain injury with a high recurrence rate. The aim of this study was to identify previously unknown MTJ components in mature animals and humans. Samples were obtained from the superficial digital flexor (SDF) muscle-tendon interface of 20 horses, and the tissue was separated through a sequential cryosectioning approach into muscle, MTJ (muscle tissue enriched in myofiber tips attached to the tendon), and tendon fractions. RT-PCR was performed for genes known to be expressed in the three tissue fractions and t-distributed stochastic neighbor embedding (t-SNE) plots were used to select the muscle, MTJ, and tendon samples from five horses for RNA sequencing. The expression of previously known and unknown genes identified through RNA sequencing was studied by immunofluorescence on human hamstring MTJ tissue. The main finding was that RNA sequencing identified the expression of a panel of 61 genes enriched at the MTJ. Of these, 48 genes were novel for the MTJ and 13 genes had been reported to be associated with the MTJ in earlier studies. The expression of known [COL22A1 (collagen XXII), NCAM (neural cell adhesion molecule), POSTN (periostin), NES (nestin), OSTN (musclin/osteocrin)] and previously undescribed [MNS1 (meiosis-specific nuclear structural protein 1), and LCT (lactase)] MTJ genes was confirmed at the protein level by immunofluorescence on tissue sections of human MTJ. In conclusion, in muscle-tendon interface tissue enriched with myofiber tips, we identified the expression of previously unknown MTJ genes representing diverse biological processes, which may be important in the maintenance of the specialized MTJ.
Subject(s)
Hamstring Muscles/metabolism , Hamstring Tendons/metabolism , Muscle Fibers, Skeletal/metabolism , Muscle Proteins/genetics , RNA, Messenger/genetics , Adult , Animals , Cell Adhesion Molecules/genetics , Cell Adhesion Molecules/metabolism , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Collagen/genetics , Collagen/metabolism , Female , Fluorescent Antibody Technique , Gene Expression Profiling , Gene Expression Regulation , Gene Ontology , Horses , Humans , Male , Molecular Sequence Annotation , Muscle Proteins/classification , Muscle Proteins/metabolism , Nestin/genetics , Nestin/metabolism , Neural Cell Adhesion Molecules/genetics , Neural Cell Adhesion Molecules/metabolism , RNA, Messenger/classification , RNA, Messenger/metabolism , Sequence Analysis, RNA , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
OBJECTIVE: To characterize muscle involvement and evaluate disease severity in patients with GNE myopathy using skeletal muscle MRI and proton magnetic resonance spectroscopy (1H-MRS). METHODS: Skeletal muscle imaging of the lower extremities was performed in 31 patients with genetically confirmed GNE myopathy, including T1-weighted and short tau inversion recovery (STIR) images, T1 and T2 mapping, and 1H-MRS. Measures evaluated included longitudinal relaxation time (T1), transverse relaxation time (T2), and 1H-MRS fat fraction (FF). Thigh muscle volume was correlated with relevant measures of strength, function, and patient-reported outcomes. RESULTS: The cohort was representative of a wide range of disease progression. Contractile thigh muscle volume ranged from 5.51% to 62.95% and correlated with thigh strength (r = 0.91), the 6-minute walk test (r = 0.82), the adult myopathy assessment tool (r = 0.83), the activities-specific balance confidence scale (r = 0.65), and the inclusion body myositis functional rating scale (r = 0.62). Four stages of muscle involvement were distinguished by qualitative (T1W and STIR images) and quantitative methods: stage I: unaffected muscle (T1 = 1,033 ± 74.2 ms, T2 = 40.0 ± 1.9 ms, FF = 7.4 ± 3.5%); stage II: STIR hyperintense muscle with minimal or no fat infiltration (T1 = 1,305 ± 147 ms, T2 = 50.2 ± 3.5 ms, FF = 27.6 ± 12.7%); stage III: fat infiltration and STIR hyperintensity (T1 = 1,209 ± 348 ms, T2 = 73.3 ± 12.6 ms, FF = 57.5 ± 10.6%); and stage IV: complete fat replacement (T1 = 318 ± 39.9 ms, T2 = 114 ± 21.2 ms, FF = 85.6 ± 4.2%). 1H-MRS showed a significant decrease in intramyocellular lipid and trimethylamines between stage I and II, suggesting altered muscle metabolism at early stages. CONCLUSION: MRI biomarkers can monitor muscle involvement and determine disease severity noninvasively in patients with GNE myopathy. CLINICALTRIALSGOV IDENTIFIER: NCT01417533.
Subject(s)
Distal Myopathies/diagnostic imaging , Lipid Metabolism , Muscle Strength , Muscle, Skeletal/diagnostic imaging , Adult , Aged , Disease Progression , Distal Myopathies/metabolism , Distal Myopathies/pathology , Distal Myopathies/physiopathology , Female , Hamstring Muscles/diagnostic imaging , Hamstring Muscles/metabolism , Hamstring Muscles/pathology , Hamstring Muscles/physiopathology , Humans , Leg , Lipids , Magnetic Resonance Imaging , Male , Middle Aged , Multienzyme Complexes/genetics , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathology , Muscle, Skeletal/physiopathology , Organ Size , Patient Reported Outcome Measures , Proton Magnetic Resonance Spectroscopy , Quadriceps Muscle/diagnostic imaging , Quadriceps Muscle/metabolism , Quadriceps Muscle/pathology , Severity of Illness Index , Thigh , Walk Test , Young AdultABSTRACT
It was hypothesized that single-nucleotide polymorphisms (SNPs) extracted from text-mined genes could be more tightly related to causal variant for each trait and that differentially weighting of this SNP panel in the GBLUP model could improve the performance of genomic prediction in cattle. Fitting two GRMs constructed by text-mined SNPs and SNPs except text-mined SNPs from 777k SNPs set (exp_777K) as different random effects showed better accuracy than fitting one GRM (Im_777K) for six traits (e.g. backfat thickness: +â 0.002, eye muscle area: + 0.014, Warner-Bratzler Shear Force of semimembranosus and longissimus dorsi: + 0.024 and + 0.068, intramuscular fat content of semimembranosus and longissimus dorsi: + 0.008 and +â 0.018). These results can suggest that attempts to incorporate text mining into genomic predictions seem valuable, and further study using text mining can be expected to present the significant results.
Subject(s)
Genome-Wide Association Study , Genome/genetics , Quantitative Trait Loci/genetics , Animals , Breeding , Cattle , Data Mining , Genomics , Genotype , Hamstring Muscles/growth & development , Hamstring Muscles/metabolism , Humans , Models, Genetic , Pedigree , Phenotype , Polymorphism, Single Nucleotide/genetics , Republic of KoreaABSTRACT
The aim of the study was to compare the transcriptomic profiles of fully differentiated skeletal muscle derived from bulls belonging to different breeds of varying performance. Microarray analyses were performed to determine the differences in the expression profiles of genes between semitendinosus muscles of 15-month-old beef-breed bulls (Limousin-LIM and Hereford-HER) and dairy-breed bulls (Holstein Friesian-HF). These analyses allowed for the identification of those genes the expression of which is similar and characteristic of fully differentiated muscle in beef breeds, but differs in skeletal muscle of a typical dairy breed. The analysis revealed 463 transcripts showing similar expression in the semitendinosus muscle of beef breeds (LIM/HER), in comparison with the dairy breed (HF). Among the identified genes, 227 were upregulated and 236 were downregulated in beef breeds. The ontological analyses revealed that the largest group of genes similarly expressed in LIM and HER was involved in the processes of protein metabolism and development of muscle organ. In beef breeds, some genes involved in protein synthesis and proteolysis showed an upregulation, including ctsd, ctsf, fhl2, fhl3, fst, sirt1, and trim63, whereas some were downregulated, including bmpr1a, bmpr2, mstn, smad2, hspa8, gsk3ß, and tgfß2. The expression of the chosen genes was confirmed by RT-qPCR technique. Thus, it can be assumed that the identified genes involved in the regulation of growth and development of muscle tissue and the processes of protein metabolism in the examined cattle breeds may be responsible for the greater gain of muscle mass in beef-breed bulls.
Subject(s)
Breeding , Hamstring Muscles/metabolism , Transcriptome/genetics , Animals , Cattle , Cell Differentiation , Male , Meat/analysisABSTRACT
Fetal sheep with placental insufficiency-induced intrauterine growth restriction (IUGR) have lower hindlimb oxygen consumption rates (OCRs), indicating depressed mitochondrial oxidative phosphorylation capacity in their skeletal muscle. We hypothesized that OCRs are lower in skeletal muscle mitochondria from IUGR fetuses, due to reduced electron transport chain (ETC) activity and lower abundances of tricarboxylic acid (TCA) cycle enzymes. IUGR sheep fetuses (n = 12) were created with mid-gestation maternal hyperthermia and compared with control fetuses (n = 12). At 132 ± 1 days of gestation, biceps femoris muscles were collected, and the mitochondria were isolated. Mitochondria from IUGR muscle have 47% lower State 3 (Complex I-dependent) OCRs than controls, whereas State 4 (proton leak) OCRs were not different between groups. Furthermore, Complex I, but not Complex II or IV, enzymatic activity was lower in IUGR fetuses compared with controls. Proteomic analysis (n = 6/group) identified 160 differentially expressed proteins between groups, with 107 upregulated and 53 downregulated mitochondria proteins in IUGR fetuses compared with controls. Although no differences were identified in ETC subunit protein abundances, abundances of key TCA cycle enzymes [isocitrate dehydrogenase (NAD+) 3 noncatalytic subunit ß (IDH3B), succinate-CoA ligase ADP-forming subunit-ß (SUCLA2), and oxoglutarate dehydrogenase (OGDH)] were lower in IUGR mitochondria. IUGR mitochondria had a greater abundance of a hypoxia-inducible protein, NADH dehydrogenase 1α subcomplex 4-like 2, which is known to incorporate into Complex I and lower Complex I-mediated NADH oxidation. Our findings show that mitochondria from IUGR skeletal muscle adapt to hypoxemia and hypoglycemia by lowering Complex I activity and TCA cycle enzyme concentrations, which together, act to lower OCR and NADH production/oxidation in IUGR skeletal muscle.
Subject(s)
Citric Acid Cycle/physiology , Electron Transport Complex I/metabolism , Fetal Growth Retardation/metabolism , Mitochondria, Muscle/metabolism , Animals , Down-Regulation , Electron Transport Complex II/metabolism , Electron Transport Complex IV/metabolism , Female , Fetal Growth Retardation/enzymology , Hamstring Muscles/enzymology , Hamstring Muscles/metabolism , Hypoglycemia/enzymology , Hypoglycemia/metabolism , Hypoxia/enzymology , Hypoxia/metabolism , Isocitrate Dehydrogenase/metabolism , Ketoglutarate Dehydrogenase Complex/metabolism , Mitochondria, Muscle/enzymology , Mitochondrial Proteins/metabolism , Muscle, Skeletal/enzymology , Muscle, Skeletal/metabolism , Oxygen Consumption , Placental Insufficiency/enzymology , Placental Insufficiency/metabolism , Pregnancy , Proteomics , Sheep , Succinate-CoA Ligases/metabolism , Up-RegulationABSTRACT
This study mainly investigated the effect of different salt concentrations (1, 3, or 5%) on triglycerides (TG) hydrolysis in muscle during salting by analyzing moisture distribution, TG hydrolysis, TG hydrolase activity, native and phosphorylated adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) protein content, lipid droplets morphology, and muscle microstructure. The results showed that increasing salt concentration could significantly decrease T21 moisture proportion and relaxation time (p < 0.05), which was more beneficial to the lipase activity. The TG hydrolase activity increased first and then decreased with the salt concentration increasing during dry-salting process, and 3% salt concentration was the point of inflection. Western blot (WB) analysis detected both ATGL, HSL and their phosphorylated proteins, which were increased with the salt content increase. The microstructure analysis showed that the lipid droplets were split into small lipid droplets with the increase of salt content, which was more conducive to the triglycerides hydrolysis.
Subject(s)
Adipose Tissue/metabolism , Hamstring Muscles/metabolism , Lipase/metabolism , Lipid Droplets/metabolism , Sodium Chloride/pharmacology , Sterol Esterase/metabolism , Triglycerides/metabolism , Adipose Tissue/drug effects , Animals , Hamstring Muscles/drug effects , Hydrolysis , Lipid Droplets/drug effects , Phosphorylation , SwineABSTRACT
Proteomics offers the opportunity to identify and quantify many proteins and to explore how they correlate and interact with each other in biological networks. This study aimed to characterize changes in the muscle proteome during the destruction, repair, and early-remodeling phases after impact trauma in male Wistar rats. Muscle tissue was collected from uninjured control rats and rats that were euthanized between 6 h and 14 days after impact injury. Muscle tissue was analyzed using unbiased, data-independent acquisition LC-MS/MS. We identified 770 reviewed proteins in the muscle tissue, 296 of which were differentially abundant between the control and injury groups (P ≤ 0.05). Around 50 proteins showed large differences (≥10-fold) or a distinct pattern of abundance after injury. These included proteins that have not been identified previously in injured muscle, such as ferritin light chain 1, fibrinogen γ-chain, fibrinogen ß-chain, osteolectin, murinoglobulin-1, T-kininogen 2, calcium-regulated heat-stable protein 1, macrophage-capping protein, retinoid-inducible serine carboxypeptidase, ADP-ribosylation factor 4, Thy-1 membrane glycoprotein, and ADP-ribosylation factor-like protein 1. Some proteins increased between 6 h and 14 days, whereas other proteins increased in a more delayed pattern at 7 days after injury. Bioinformatic analysis revealed that various biological processes, including regulation of blood coagulation, fibrinolysis, regulation of wound healing, tissue regeneration, acute inflammatory response, and negative regulation of the immune effector process, were enriched in injured muscle tissue. This study advances the understanding of early muscle healing after muscle injury and lays a foundation for future mechanistic studies on interventions to treat muscle injury.
Subject(s)
Blood Coagulation , Fibrinolysis , Inflammation , Muscle, Skeletal/metabolism , Regeneration , Wound Healing , Wounds, Nonpenetrating/metabolism , Animals , Chromatography, Liquid , Computational Biology , Gracilis Muscle/injuries , Gracilis Muscle/metabolism , Hamstring Muscles/injuries , Hamstring Muscles/metabolism , Kinetics , Male , Muscle, Skeletal/injuries , Muscle, Skeletal/pathology , Necrosis , Proteome/metabolism , Proteomics , Rats , Tandem Mass Spectrometry , Wounds, Nonpenetrating/pathologyABSTRACT
We have previously found that in utero exposure to excess maternal cortisol (1 mg/kg/day) in late gestation increases the incidence of stillbirth during labor and produces fetal bradycardia at birth. In the interventricular septum, mitochondrial DNA (mt-DNA) was decreased, and transcriptomics and metabolomics were consistent with altered mitochondrial metabolism. The present study uses transcriptomics to model effects of increased maternal cortisol on fetal biceps femoris. Transcriptomic modeling revealed that pathways related to mitochondrial metabolism were downregulated, whereas pathways for regulation of reactive oxygen species and activation of the apoptotic cascade were upregulated. Mt-DNA and the protein levels of cytochrome C were significantly decreased in the biceps femoris. RT-PCR validation of the pathways confirmed a significant decrease in SLC2A4 mRNA levels and a significant increase in PDK4, TXNIP, ANGPTL4 mRNA levels, suggesting that insulin sensitivity of the biceps femoris muscle may be reduced in cortisol offspring. We also tested for changes in gene expression in diaphragm by rt-PCR. PDK4, TXNIP, and ANGPTL4 mRNA were also increased in the diaphragm, but SLC2A4, cytochrome C protein, and mt-DNA were unchanged. Comparison of the change in gene expression in biceps femoris to that in cardiac interventricular septum and left ventricle showed few common genes and little overlap in specific metabolic or signaling pathways, despite reduction in mt-DNA in both heart and biceps femoris. Our results suggest that glucocorticoid exposure alters expression of nuclear genes important to mitochondrial activity and oxidative stress in both cardiac and skeletal muscle tissues, but that these effects are tissue-specific.
Subject(s)
Fetal Development/drug effects , Fetal Development/genetics , Fetal Heart/drug effects , Hamstring Muscles/metabolism , Hydrocortisone/pharmacology , Myocardium/metabolism , Transcriptome , Animals , Cytochromes c/metabolism , DNA, Mitochondrial/metabolism , Female , Fetal Heart/metabolism , Gene Expression/drug effects , Glucose Transporter Type 4/metabolism , Mitochondria/drug effects , Mitochondria/metabolism , Oxidative Stress/drug effects , Pregnancy , Sheep , Signal Transduction/drug effectsABSTRACT
The objective of this study was to determine the impact of quality grade and steak location on color stability of semitendinosus (ST) steaks during a 9-d refrigerated study. Twenty-one ST muscles (12 Choice and 9 Select) were purchased from a commercial beef packing plant and fabricated into twelve 2.54-cm thick steaks per muscle. Steaks 1, 6, and 12 were designated for immunohistochemistry while remaining steak locations of proximal (steaks 2 to 4), middle (5 to 8), and distal (9 to 11) were randomly assigned to 0, 4, or 9 d of simulated retail display. Surface color attributes of day-9 steaks were recorded daily by a visual color panel and spectrophotometer. On days 0, 4, and 9 of display, steaks were analyzed for metmyoglobin reducing ability (MRA) and oxygen consumption (OC). Grade × day of display (DOD) interactions were detected for L*, a*, surface oxymyoglobin (OMb) and metmyoglobin (MMb) percentages, and visual panel surface redness and discoloration scores (P ≤ 0.02); however, no Grade × DOD interactions were observed for MRA or OC (P > 0.17). There were location main effect (LOC) × DOD interactions for L*, a*, surface MMb, visual panel surface redness and discoloration, and MRA (P ≤ 0.02). Distal steaks had lower L* values compared with the other locations (P < 0.01), which coincided with steaks being rated visually darker red (P < 0.01). Proximal steaks had greater a* values and had less surface discoloration than middle steaks (P < 0.05), which had an increased percentage of surface MMb (P ≤ 0.04). Distal and proximal steaks had increased MRA compared with middle steaks on days 0 and 4 (P < 0.05), and distal steaks had greater OC than the other locations throughout display (P < 0.01). There were fewer type I fibers at the proximal end with a greater percentage located at the middle and distal ends, and an increased percentage of type IIX fibers at the middle and proximal locations (P ≤ 0.01). Less type IIA fibers were detected at the middle LOC compared with the other two locations (P < 0.10). Larger type I, IIA, and IIX fibers were located at the proximal and middle locations compared with the distal LOC (P < 0.01). ST color and color-stability characteristics were influenced by DOD and LOC, which may partially be explained by differences in fiber types among locations.
Subject(s)
Cattle/metabolism , Oxygen Consumption , Red Meat/standards , Animals , Color , Hamstring Muscles/metabolism , Metmyoglobin/analysis , Metmyoglobin/metabolism , Muscle Fibers, Skeletal/metabolism , Myoglobin/analysis , Myoglobin/metabolism , NAD/analysis , NAD/metabolism , Random Allocation , Red Meat/analysisABSTRACT
Effect of sous-vide processing was evaluated on in-vitro simulated protein digestion, myofibrillar protein profile, lipid oxidation and physicochemical properties of cold-boned beef Semitendinosus. Sous-vide processed [T1, 60⯰C (4.5â¯h); T2, 60⯰C (10â¯h)] samples along with control beef (cooked at 80⯰C until a core temperature of 75⯰C was attained) were subjected to in-vitro simulated gastrointestinal protein digestion. Samples collected at 0, 30 and 60â¯min of gastric digestion and 120 and 180â¯min of intestinal digestion were analysed for protein profile (SDS-PAGE), protein digestibility (%), soluble protein (%), free amino acid analysis and mineral profile. A significant (Pâ¯<â¯0.05) decrease was observed in shear force (N) and cooking loss (%) of sous-vide processed samples in comparison to control whereas a significant (Pâ¯<â¯0.05) increase was observed in colour (L*, a*, b*). A significant (Pâ¯<â¯0.05) increase was also observed in protein digestibility (%), soluble protein (%) and release of free amino acids and minerals from sous-vide processed samples during in-vitro digestion. By modifying the protein profile of the meat, sous-vide processing had a positive influence on in-vitro digestion kinetics that led to a greater and faster digestion of proteins during simulated digestion.
Subject(s)
Cooking/methods , Food Quality , Hamstring Muscles/metabolism , Proteolysis , Red Meat/analysis , Animals , Cattle , In Vitro TechniquesABSTRACT
In order to investigate the role of the Notch signaling pathway in skeletal muscle fibrosis after nerve injury, 60 Sprague-Dawley rats were selected and divided randomly into a control and two experimental groups. Group A served as controls without any treatment. Rats in groups B were injected intraperitoneally with 0.2 mL PBS and those in group C were injected intraperitoneally with 0.2 mL PBS+100 µmol/L, 0.2 mL N-[N-(3,5-difluorophenacetyl)-l-alanyl]- S-phenylglycine t-butyl ester (DAPT, a gamma-secretase inhibitor that suppresses Notch signaling) respectively, on postoperative days 1, 3, 7, 10, and 14 in a model of denervation-induced skeletal muscle fibrosis by right sciatic nerve transection. Five rats from each group were euthanized on postoperative days 1, 7, 14, and 28 to collect the right gastrocnemii, and hematoxylin and eosin (HE) staining, immunohistochemistry test, real-time PCR, and Western blotting were performed to assess connective tissue hyperplasia and fibroblast density as well as expression of Notch 1, Jagged 1, and Notch downstream molecules Hes 1 and collagen I (COL I) on day 28. There was no significant difference in HE-stained fibroblast density between group B and C on postoperative day 1. However, fibroblast density was significantly higher in group B than in group C on postoperative days 7, 14, and 28. Notch 1, Jagged 1, Hes 1, and COL I proteins in the gastrocnemius were expressed at very low levels in group A but at high levels in group B. Expression levels of these proteins were significantly lower in group C than in group B (P<0.05), but they were higher in group C than in group A (P<0.05) on postoperative day 28. We are led to conclude that locking the Notch signaling pathway inhibits fibrosis progression of denervated skeletal muscle. Thus, it may be a new approach for treatment of fibrosis of denervated skeletal muscle.
Subject(s)
Amyloid Precursor Protein Secretases/genetics , Dipeptides/pharmacology , Enzyme Inhibitors/pharmacology , Fibroblasts/metabolism , Receptor, Notch1/genetics , Sciatic Nerve/drug effects , Amyloid Precursor Protein Secretases/antagonists & inhibitors , Amyloid Precursor Protein Secretases/metabolism , Animals , Collagen Type I/genetics , Collagen Type I/metabolism , Fibroblasts/drug effects , Fibroblasts/pathology , Fibrosis/prevention & control , Gene Expression Regulation , Hamstring Muscles/drug effects , Hamstring Muscles/innervation , Hamstring Muscles/metabolism , Injections, Intraperitoneal , Jagged-1 Protein/genetics , Jagged-1 Protein/metabolism , Male , Muscle Denervation/methods , Muscle, Skeletal/drug effects , Muscle, Skeletal/innervation , Muscle, Skeletal/metabolism , Rats , Rats, Sprague-Dawley , Receptor, Notch1/antagonists & inhibitors , Receptor, Notch1/metabolism , Sciatic Nerve/injuries , Sciatic Nerve/metabolism , Signal Transduction , Transcription Factor HES-1/genetics , Transcription Factor HES-1/metabolismABSTRACT
BACKGROUND: Korean Hanwoo cattle are known for their high meat quality, especially their high intramuscular fat compared to most other cattle breeds. Different muscles have very different meat quality traits and a study of the myogenic process in satellite cells can help us better understand the genes and pathways that regulate this process and how muscles differentiate. RESULTS: Cell cultures of Longissimus dorsi muscle differentiated from myoblast into multinucleated myotubes faster than semimembranosus. Time-series RNA-seq identified a total of 13 differentially expressed genes between the two muscles during their development. These genes seem to be involved in determining muscle lineage development and appear to modulate the expression of myogenic regulatory factors (mainly MYOD and MYF5) during differentiation of satellite cells into multinucleate myotubes. Gene ontology enriched terms were consistent with the morphological changes observed in the histology. Most of the over-represented terms and genes expressed during myoblast differentiation were similar regardless of muscle type which indicates a highly conserved myogenic process albeit the rates of differentiation being different. There were more differences in the enriched GO terms during the end of proliferation compared to myoblast differentiation. CONCLUSIONS: The use of satellite cells from newborn Hanwoo calves appears to be a good model to study embryonic myogenesis in muscle. Our findings provide evidence that the differential expression of HOXB2, HOXB4, HOXB9, HOXC8, FOXD1, IGFN1, ZIC2, ZIC4, HOXA11, HOXC11, PITX1, SIM2 and TBX4 genes could be involved in the differentiation of Longissimus dorsi and Semimembranosus muscles. These genes seem to modulate the muscle fate of the satellite cells during myogenesis through a differential expression profile that also controls the expression of some myogenic regulatory factors (MYOD and MYF5). The number of differentially expressed genes across time was unsurprisingly large. In relation to the baseline day 0, there were 631, 155, 175, 519 and 586 DE genes in LD, while in SM we found 204, 0, 615, 761 and 1154 DE genes at days 1, 2, 4, 7 and 14 respectively.
Subject(s)
Back Muscles/cytology , Cattle/genetics , Hamstring Muscles/cytology , Satellite Cells, Skeletal Muscle/cytology , Satellite Cells, Skeletal Muscle/metabolism , Animals , Back Muscles/metabolism , Cattle/anatomy & histology , Cell Differentiation , Cells, Cultured , Gene Expression Profiling , Genome , Hamstring Muscles/metabolism , Sequence Alignment , Sequence Analysis, RNAABSTRACT
BACKGROUND: Late-onset multiple acyl-coA dehydrogenase deficiency (MADD) is an autosomal recessive inherited metabolic disorder. It is still unclear about the muscle magnetic resonance image (MRI) pattern of the distal lower limb pre- and post-treatment in patients with late-onset MADD. This study described the clinical and genetic findings in a cohort of patients with late-onset MADD, and aimed to characterize the MRI pattern of the lower limbs. METHODS: Clinical data were retrospectively collected from clinic centers of Peking University People's Hospital between February 2014 and February 2018. Muscle biopsy, blood acylcarnitines, and urine organic acids profiles, and genetic analysis were conducted to establish the diagnosis of MADD in 25 patients. Muscle MRI of the thigh and leg were performed in all patients before treatment. Eight patients received MRI re-examinations after treatment. RESULTS: All patients presented with muscle weakness or exercise intolerance associated with variants in the electron transfer flavoprotein dehydrogenase gene. Muscle MRI showed a sign of both edema-like change and fat infiltration selectively involving in the soleus (SO) but sparing of the gastrocnemius (GA) in the leg. Similar sign of selective involvement of the biceps femoris longus (BFL) but sparing of the semitendinosus (ST) was observed in the thigh. The sensitivity and specificity of the combination of either "SO+/GA-" sign or "BFL+/ST-" sign for the diagnosis of late-onset MADD were 80.0% and 83.5%, respectively. Logistic regression model supported the findings. The edema-like change in the SO and BFL muscles were quickly recovered at 1 month after treatment, and the clinical symptom was also relieved. CONCLUSIONS: This study expands the clinical and genetic spectrums of late-onset MADD. Muscle MRI shows a distinct pattern in the lower limb of patients with late-onset MADD. The dynamic change of edema-like change in the affected muscles might be a potential biomarker of treatment response.