Proteomic Changes in Sarcoplasmic and Myofibrillar Proteins Associated with Color Stability of Ovine Muscle during Post-Mortem Storage.
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ABSTRACT:
SUBMITTER: Gao X
PROVIDER: S-EPMC8700786 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:This study investigated the effect of different cattle management strategies at farm (Intensive vs. Extensive) and during transport and lairage (mixing vs. non-mixing with unfamiliar animals) on the myofibrillar subproteome of Longissimus thoracis et lumborum (LTL) muscle of "Asturiana de los Valles" yearling bulls. It further aimed to study the relationships with beef quality traits including pH, color, and tenderness evaluated by Warner-Bratzler shear force (WBSF). Thus, comparative proteomics of the myofibrillar fraction along meat maturation (from 2 h to 14 days post-mortem) and different quality traits were analyzed. A total of 23 protein fragments corresponding to 21 unique proteins showed significant differences among the treatments (p < 0.05) due to any of the factors considered (Farm, Transport and Lairage, and post-mortem time ageing). The proteins belong to several biological pathways including three structural proteins (MYBPC2, TNNT3, and MYL1) and one metabolic enzyme (ALDOA) that were affected by both Farm and Transport/Lairage factors. ACTA1, LDB3, and FHL2 were affected by Farm factors, while TNNI2 and MYLPF (structural proteins), PKM (metabolic enzyme), and HSPB1 (small Heat shock protein) were affected by Transport/Lairage factors. Several correlations were found between the changing proteins (PKM, ALDOA, TNNI2, TNNT3, ACTA1, MYL1, and CRYAB) and color and tenderness beef quality traits, indicating their importance in the determination of meat quality and their possible use as putative biomarkers.
| S-EPMC8700955 | biostudies-literature
Project description:The composition and function of fragmented sarcoplasmic reticulum from pig skeletal muscle was examined in the period immediately post mortem. Muscle was defined as being either slowly glycolysing or rapidly glycolysing on the basis of colour, pH and concentrations of glycogen and lactate. The microsomal fraction was separated on a discontinuous gradient of 35, 40 and 45% (w/v) sucrose into heavy and intermediate fractions which sedimented to the interfaces, and a light fraction which remained on the surface of the 35%-sucrose layer. The sarcoplasmic reticulum from rapidly glycolysing muscle had a lower buoyant density than had that from slowly glycolysing muscle. This was reflected in the consistent lack of material in the heavy fraction and a greater proportion in the light fraction. The latter material had significantly lower ratios (w/w) of protein to phospholipid (2.3:1 versus 3.8:1) and of protein to cholesterol (10.4:1 versus 15.6:1). There were no gross differences in phospholipid content or in fatty acid composition of individual phospholipid classes in the membranes from the two types of muscle. Analysis of membrane proteins by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis showed that ATPase (adenosine triphosphatase) was a major component of each fraction and that its contribution to the total protein content of the membrane was greater in rapidly glycolysing muscle, suggesting a loss of non-ATPase proteins. The two fractions of sarcoplasmic reticulum prepared from rapidly glycolysing muscle had approximately one-third the normal activities of Ca(2+) binding and Ca(2+) uptake in the presence of ATP and one-half the passive Ca(2+)-binding capacity in the absence of ATP of the fractions from slowly glycolysing muscle. However, the (Ca(2+)+Mg(2+))-stimulated ATPase activities were similar. Efflux from actively loaded vesicles, after the addition of EDTA, consisted of a rapid and a slow phase. Vesicles from rapidly glycolysing muscle lost 60% of associated Ca(2+) (approx. 0.10mumol of Ca(2+)/mg of protein) during the rapid phase, compared with 30% (approx. 0.17mumol of Ca(2+)/mg of protein) in those from slowly glycolysing muscle. The efflux rate during the slower phase was comparable in both types of vesicles. Analysis of the temperature-dependence of (Ca(2+)+Mg(2+))-stimulated ATPase activity revealed that a high-activation-energy process operating in the temperature range 31-45 degrees C in the intermediate and light fractions from slowly glycolysing muscle was not apparent in vesicles from rapidly glycolysing muscle. Conditions that result in the prolonged activation of glycogenolysis in pig muscle post mortem primarily affect the protein components of the sarcoplasmic-reticular membrane, giving rise to a loss of loosely associated proteins. The function of the membranes observed under these conditions does not appear to be due to enhanced permeability of the membrane to Ca(2+) and may be the result of a defect in the transport of Ca(2+) into the vesicles.
| S-EPMC1165021 | biostudies-other
Project description:BackgroundRecovering high quality intact RNA from post-mortem tissue is of major concern for gene expression studies in animals and humans. Since the availability of post-mortem tissue is often associated with substantial delay, it is important that we understand the temporal variation in the stability of total RNA and of individual gene transcripts so as to be able to appropriately interpret the data generated from such studies. Hence, the objective of this experiment was to qualitatively and quantitatively assess the integrity of total and messenger RNA extracted from bovine skeletal muscle, subcutaneous adipose tissue and liver stored at 4 degrees C at a range of time points up to 22 days post-mortem. These conditions were designed to mimic the environment prevailing during the transport of beef from the abattoir to retail outlets.ResultsThe 28S and 18S rRNA molecules of total RNA were intact for up to 24 h post-mortem in liver and adipose tissues and up to 8 days post-mortem in skeletal muscle. The mRNA of housekeeping genes (GAPDH and ACTB) and two diet-related genes (RBP5 and SCD) were detectable up to 22 days post-mortem in skeletal muscle. While the mRNA stability of the two housekeeping genes was different in skeletal muscle and liver, they were similar to each other in adipose tissue. After 22 days post-mortem, the relative abundance of RBP5 gene was increased in skeletal muscle and in adipose tissue and decreased in liver. During this period, the relative abundance of SCD gene also increased in skeletal muscle whereas it decreased in both adipose tissue and liver.ConclusionStability of RNA in three tissues (skeletal muscle, subcutaneous adipose tissue and liver) subjected to long-term post-mortem storage at refrigeration temperature indicated that skeletal muscle can be a suitable tissue for recovering biologically useful RNA for gene expression studies even if the tissue is subjected to post-mortem storage for weeks, whereas adipose tissue and liver should be processed within 24 hours post-mortem.
| S-EPMC2234424 | biostudies-literature
Project description:BackgroundEpigenetic (including DNA and histone) modifications occur in a variety of neurological disorders. If epigenetic features of brain autopsy material are to be studied, it is critical to understand the post-mortem stability of the modifications.MethodsPig and mouse brain tissue were formalin-fixed and paraffin-embedded, or frozen after post-mortem delays of 0, 24, 48, and 72?h. Epigenetic modifications frequently reported in the literature were studied by DNA agarose gel electrophoresis, DNA methylation enzyme-linked immunosorbent assays, Western blotting, and immunohistochemistry. We constructed a tissue microarray of human neocortex samples with devitalization or death to fixation times ranging from <?60?min to 5?days.ResultsIn pig and mouse brain tissue, we found that DNA cytosine modifications (5mC, 5hmC, 5fC, and 5caC) were stable for ??72?h post-mortem. Histone methylation was generally stable for ??48?h (H3K9me2/K9me3, H3K27me2, H3K36me3) or ??72?h post-mortem (H3K4me3, H3K27me3). Histone acetylation was generally less stable. The levels of H3K9ac, H3K27ac, H4K5ac, H4K12ac, and H4K16ac declined as early as ??24?h post-mortem, while the levels of H3K14ac did not change at ??48?h. Immunohistochemistry showed that histone acetylation loss occurred primarily in the nuclei of large neurons, while immunoreactivity in glial cell nuclei was relatively unchanged. In the human brain tissue array, immunoreactivity for DNA cytosine modifications and histone methylation was stable, while subtle changes were apparent in histone acetylation at 4 to 5?days post-mortem.ConclusionWe conclude that global epigenetic studies on human post-mortem brain tissue are feasible, but great caution is needed for selection of post-mortem delay matched controls if histone acetylation is of interest.
| S-EPMC6330433 | biostudies-literature
Project description:Two dimensional Difference Gel Electrophoresis (2-D DIGE) and mass spectrometry were applied to investigate the changes in metabolic proteins that occur over a seven day (day 1, 3 and 7) post mortem ageing period in porcine centrifugal exudate from divergent meat quality phenotypes. The objectives of the research were to enhance our understanding of the phenotype (water holding capacity) and search for biomarkers of this economically significant pork quality attribute. Major changes in protein abundance across nine phenotype-by-time conditions were observed. Proteomic patterns were dominated by post mortem ageing timepoint. Using a machine learning algorithm (l1-regularized logistic regression), a model was derived with the ability to discriminate between high drip and low drip phenotypes using a subset of 25 proteins with an accuracy of 63%. Models discriminating between divergent phenotypes with accuracy of 72% and 73% were also derived comparing respectively, high drip plus intermediate phenotype (considered as one phenotype) versus low drip and comparing low drip plus intermediate phenotype (considered as one phenotype) versus high drip. In all comparisons, the general classes of discriminatory proteins identified include metabolic enzymes, stress response, transport and structural proteins. In this research we have enhanced our understanding of the protein related processes underpinning this phenotype and provided strong data to work toward development of protein biomarkers for water holding capacity.
| S-EPMC4780776 | biostudies-literature
Project description:Proteomic characterization of human brain tissue is increasingly utilized to identify potential novel biomarkers and drug targets for a variety of neurological diseases. In whole-tissue studies, results may be driven by changes in the proportion of the largest and most abundant organelles or tissue cell-type composition. Spatial proteomics approaches enhance our knowledge of disease mechanisms and changing signalling pathways at the subcellular level by taking into account the importance of cellular localization, which critically influences protein function. Density gradient-based ultracentrifugation methods allow for subcellular fractionation and have been utilized in cell lines, mouse and human brain tissue to quantify thousands of proteins in specific enriched organelles such as the pre- and post-synapse. Serial ultracentrifugation methods allow for the analysis of multiple cellular organelles from the same biological sample, and to our knowledge have not been previously applied to frozen post-mortem human brain tissue. The use of frozen human tissue for tissue fractionation faces two major challenges, the post-mortem interval, during which proteins may leach from their usual location into the cytosol, and freezing, which results in membrane breakdown. Despite these challenges, in this proof-of-concept study, we show that the majority of proteins segregate reproducibly into crude density-based centrifugation fractions, that the fractions are enriched for the appropriate organellar markers and that significant differences in protein localization can be observed between tissue from individuals with Alzheimer's disease and control individuals.
| S-EPMC9123841 | biostudies-literature
Project description:The aim of this study was to compare color shift, color stability, and post-polishing surface roughness of esthetic restorative materials. Twenty-five disc-shaped specimens (10 mm in diameter and 2 mm in thickness) from five esthetic resin materials (Z250XT, IPS Empress-Direct, G-ænial, Vit-l-escence, and Ceram.X) were fabricated. Color shift before and immediately after light-curing (?E0) then color stability after immersion in tea, coffee, berry juice, and distilled water were measured using a spectrophotometer. Color changes were measured after 2 (?E2), 4 (?E4), 6 (?E6), and 8 weeks (?E8). Surface roughness values (Rq) were determined using an optical profilometer after polishing using a rotary polishing system. Data were analyzed using ANOVA and multiple comparison methods at 0.05 significance level. There were no significant differences between the tested materials regarding color shift except between Vit-l-escence and Ceram.X (p = 0.033). There was no significant difference between coffee and tea solutions (p = 1.0) and between berry juice and distilled water (p = 0.15). There was no difference between the tested materials regarding Rq (p = 0.057). Ceram.X was associated with the lowest ?E8 values overall. Tested materials were comparable regarding color shift and post-polishing surface roughness. Susceptibility to staining was dependent on the material.
| S-EPMC7143460 | biostudies-literature
Project description:We set up a pilot study using Affymetrix Gene Chip® Porcine Genome Arrays to evaluate the impact of time lags from death on gene expression profiling of porcine skeletal muscle at four post mortem time points (up to 24 hrs) during the routine processing of fresh tights
2011-01-19 | GSE26701 | GEO
Project description:In this study the polymorphism of myofibrillar proteins and the Ca2+-uptake activity of sarcoplasmic reticulum were analysed in single fibres from human skeletal muscles. Two populations of histochemically identified type-I fibres were found differing in the number of light-chain isoforms of the constituent myosin, whereas the pattern of light chains of fast myosin of type-IIA and type-IIB fibres was indistinguishable. Regulatory proteins, troponin and tropomyosin, and other myofibrillar proteins, such as M- and C-proteins, showed specific isoforms in type-I and type-II fibres. Furthermore, tropomyosin presented different stoichiometries of the alpha- and beta-subunits between the two types of fibres. Sarcoplasmic-reticulum volume, as indicated by the maximum capacity for calcium oxalate accumulation, was almost identical in type-I and type-II fibres, whereas the rate of Ca2+ transport was twice as high in type-II as compared with type-I fibres. It is concluded that, in normal human muscle fibres, there is a tight segregation of fast and slow isoforms of myofibrillar proteins that is very well co-ordinated with the relaxing activity of the sarcoplasmic reticulum. These findings may thus represent a molecular correlation with the differences of the twitch-contraction time between fast and slow human motor units. This tight segregation is partially lost in the muscle fibres of elderly individuals.
| S-EPMC1144416 | biostudies-other
Project description:BackgroundMeat quality is a complex trait influenced by a range of factors with post mortem biochemical processes highly influential in defining ultimate quality. High resolution two-dimensional DIfference Gel Electrophoresis (2-D DIGE) and Western blot were applied to study the influence of post mortem meat ageing on the proteome of pork muscle. Exudate collected from the muscle following centrifugation was analysed at three timepoints representing a seven day meat ageing period.ResultsThe intensity of 136 spots varied significantly (p < 0.05) across this post mortem period and 40 spots were identified using mass spectrometry. The main functional categories represented were metabolic proteins, stress-related proteins, transport and structural proteins. Metabolic and structural proteins were generally observed to increase in abundance post mortem and many likely represent the accumulation of the degradation products of proteolytic enzyme activity. In contrast, stress-related proteins broadly decreased in abundance across the ageing period. Stress response proteins have protective roles in maintaining cellular integrity and a decline in their abundance over time may correlate with a reduction in cellular integrity and the onset of meat ageing. Since cellular conditions alter with muscle ageing, changes in solubility may also contribute to observed abundance profiles.ConclusionsMuscle exudate provided valuable information about the pathways and processes underlying the post mortem ageing period, highlighting the importance of post mortem modification of proteins and their interaction for the development of meat quality traits.
| S-EPMC3639075 | biostudies-literature