Cluster analysis application identifies muscle characteristics of importance for beef tenderness.

BACKGROUND: An important controversy in the relationship between beef tenderness and muscle characteristics including biochemical traits exists among meat researchers. The aim of this study is to explain variability in meat tenderness using muscle characteristics and biochemical traits available in the Integrated and Functional Biology of Beef (BIF-Beef) database. The BIF-Beef data warehouse contains characteristic measurements from animal, muscle, carcass, and meat quality derived from numerous experiments. We created three classes for tenderness (high, medium, and low) based on trained taste panel tenderness scores of all meat samples consumed (4,366 observations from 40 different experiments). For each tenderness class, the corresponding means for the mechanical characteristics, muscle fibre type, collagen content, and biochemical traits which may influence tenderness of the muscles were calculated. RESULTS: Our results indicated that lower shear force values were associated with more tender meat. In addition, muscles in the highest tenderness cluster had the lowest total and insoluble collagen contents, the highest mitochondrial enzyme activity (isocitrate dehydrogenase), the highest proportion of slow oxidative muscle fibres, the lowest proportion of fast-glycolytic muscle fibres, and the lowest average muscle fibre cross-sectional area. Results were confirmed by correlation analyses, and differences between muscle types in terms of biochemical characteristics and tenderness score were evidenced by Principal Component Analysis (PCA). When the cluster analysis was repeated using only muscle samples from m. Longissimus thoracis (LT), the results were similar; only contrasting previous results by maintaining a relatively constant fibre-type composition between all three tenderness classes. CONCLUSION: Our results show that increased meat tenderness is related to lower shear forces, lower insoluble collagen and total collagen content, lower cross-sectional area of fibres, and an overall fibre type composition displaying more oxidative fibres than glycolytic fibres.

Protocol for high-resolution electrophoresis separation of myosin heavy chain isoforms in bovine skeletal muscle.

In this short communication we describe a specific protocol for SDS-PAGE separation of adult bovine myosin heavy-chain (MyHC) isoforms. The conditions defined in this protocol allow a good separation with a good reproducibility of the four MyHC isoforms (MyHC I, IIa, IIx, IIb) identified in adult skeletal muscle of this species. This procedure uses mini-gel electrophoresis system and does not involve preparation of gradient separating gels. In addition, this protocol can also be applied to the electrophoretic separation of ovine and camel MyHC isoforms.

Specific fibre composition and metabolism of the rectus abdominis muscle of bovine Charolais cattle.

BACKGROUND: An important variability of contractile and metabolic properties between muscles has been highlighted. In the literature, the majority of studies on beef sensorial quality concerns M. longissimus thoracis. M. rectus abdominis (RA) is easy to sample without huge carcass depreciation and may appear as an alternative to M. longissimus thoracis for fast and routine physicochemical analysis. It was considered interesting to assess the muscle fibres of M. rectus abdominis in comparison with M. longissimus thoracis (LT) and M. triceps brachii (TB) on the basis of metabolic and contractile properties, area and myosin heavy chain isoforms (MyHC) proportions. Immuno-histochemical, histochemical, histological and enzymological techniques were used. This research concerned two populations of Charolais cattle: RA was compared to TB in a population of 19 steers while RA was compared to LT in a population of 153 heifers. RESULTS: RA muscle had higher mean fibre areas (3350 microm(2) vs 2142 to 2639 microm(2)) than the two other muscles. In RA muscle, the slow-oxidative fibres were the largest (3957 microm(2)) and the fast-glycolytic the smallest (2868 microm(2)). The reverse was observed in TB muscle (1725 and 2436 microm(2) respectively). In RA muscle, the distinction between fast-oxidative-glycolytic and fast-glycolytic fibres appeared difficult or impossible to establish, unlike in the other muscles. Consequently the classification based on ATPase and SDH activities seemed inappropriate, since the FOG fibres presented rather low SDH activity in this muscle in comparison to the other muscles of the carcass. RA muscle had a higher proportion of I fibres than TB and LT muscles, balanced by a lower proportion either of IIX fibres (in comparison to TB muscle) or of IIA fibres (in comparison to LT muscle). However, both oxidative and glycolytic enzyme activities were lower in RA than in TB muscle, although the LDH/ICDH ratio was higher in RA muscle (522 vs 340). Oxidative enzyme activities were higher in RA than in LT muscle, whereas glycolytic enzyme activity was lower. In RA muscle, contractile and metabolic properties appeared to be less well-correlated than in the two other muscles. CONCLUSIONS: RA muscle has some particularities in comparison to the LT and TB muscles, especially concerning the unusual large cross-section surface of SO fibres and the very low oxidative activity of intermediate IIA fibres.

In vivo proteome dynamics during early bovine myogenesis.

Myogenesis is a complex process of which the underlying mechanisms are conserved between species, including birds and mammals. Despite a good understanding of the stages of myogenesis, many of the mechanisms involved in the regulation of proliferation of the successive myoblast generations, the cellular transitions cell proliferation/alignment of myoblasts/fusion of myoblasts into myotubes/differentiation of myofibres and the control of total myofibre number still remain unknown. An in vivo proteomic analysis of the semitendinosus muscle from Charolais foetuses, at three specific stages of myogenesis (60, 110 and 180 days postconception), was conducted using 2-DE and MS. Expression profiles of more than 170 proteins were revealed and analysed using two way hierarchical clustering and statistical analysis. Our studies identify, for the first time, distinct proteins of varied biological functions and protein clusters with myogenic processes, such as the control of cell cycle activity and apoptosis, the establishment of cellular metabolism and muscle contractile properties and muscle cell reorganisation. These results are of fundamental interest to the field of myogenesis in general, and more specifically to the control of muscle development in meat producing animals.

Biochemical and transcriptomic analyses of two bovine skeletal muscles in Charolais bulls divergently selected for muscle growth.

This work aimed to investigate the consequences of muscle growth selection on muscle characteristics. An oxidative muscle (Rectus abdominis, RA) and a glycolytic one (Semitendinosus, ST) were studied in two groups of six extreme young Charolais bulls of high or low muscle growth. Mitochondrial activity was lower in muscles of bulls with high muscle growth. Transcriptomic studies allowed the identification of putatively differentially expressed genes. The differential expression between genetic types of two genes in RA (a heat shock protein and a thyroid receptor interacting protein) and of seven genes in ST (including LEU5, tropomyosin 2, and sarcosin) was confirmed by different statistical approaches or Northern blot analysis, as well as the differential expression of five genes (including PSMD4 and DPM synthase) between RA and ST. Both biochemical and transcriptomic results indicate that selection on muscle growth potential is associated with reduced slow-oxidative muscle characteristics. Further studies are required to understand the physiological importance of genes whose expression is changed by selection.

Relationships between muscle growth potential, intramuscular fat content and different indicators of muscle fibre types in young Charolais bulls.

Genetic selection in favor of muscle growth at the expense of fat should affect characteristics of muscles, and therefore beef quality. This study was conducted with two extreme groups of six animals selected among 64 Charolais young bulls ranked according to their genetic potential for muscle growth. Muscle characteristics were assessed in Rectus abdominis (RA, slow oxidative) and Semitendinosus (ST, fast glycolytic) muscles. Intramuscular fat content and proportions of myosin heavy chains I (slow) and IIA (fast oxido-glycolytic) and certain indicators of oxidative metabolism (activities of citrate synthase (CS), isocitrate dehydrogenase and cytochrome-c oxidase (COX); expression of H-fatty acid binding protein (FABP)) were higher in RA than in ST muscle. Genetic selection for muscle growth reduced intramuscular fat content and the activities of some oxidative metabolism indicators (namely CS, COX only). The positive correlation between muscle triacylglycerol content and A-FABP messenger RNA level (a marker of adipocyte differentiation) (r = 0.53, P < 0.05) suggests that A-FABP may be a good marker of the ability of bovines to deposit intramuscular fat. In conclusion, the metabolic muscle characteristics which respond to the selection process in favor of muscle growth clearly differ from the muscle characteristics which allow muscle types to be differentiated.

Meta-analysis of the comparison of the metabolic and contractile characteristics of two bovine muscles: longissimus thoracis and semitendinosus.

This study used the BIF-Beef data warehouse to determine whether semitendinosus (ST) was a muscle with a faster contraction speed and more glycolytic than longissimus thoracis (LT), regardless of the sex and breed of animals. With more than 500 animals from 7 breeds, we confirmed that LT was more oxidative than ST in males and females, but not in steers, and in all the breeds studied except Montbéliard. The LT had more slow oxidative (SO) and fewer fast oxido-glycolytic (FOG) and fast-glycolytic (FG) muscle fibres than the ST muscle, regardless of sex, in all breeds except Montbéliard and Holstein. SO proportion and the oxidative activity were negatively correlated to FG proportion and to the glycolytic activity. Similarly, FOG proportion was positively correlated to the glycolytic activity and negatively to FG proportion. However, these relationships are not consistent across sexes and breeds. In conclusion, differences in muscle types may be affected by sex or breed but to a moderate extent only.

Functional analysis of beef tenderness.

Meat tenderness represents a complex assembly of different cellular pathways. As a consequence, genomics studies have revealed many different proteins considered as tenderness markers. So it is difficult to have an overview of tenderness in terms of cellular pathways. In this work cellular pathways of tenderness were analyzed, an interactome of 330 proteins was designed, and explanations of tenderization processes were proposed.

Myogenesis is delayed in bovine fetal clones.

We have recently reported that maturation of the skeletal muscle is delayed in cloned calves during their first year postnatally. This delay could originate from perturbations in fetal myogenesis. The aim of this study was to evaluate the developmental characteristics of muscle in clones versus animals derived from conventional reproduction. We have characterized the anatomical and biochemical properties of the Semitendinosus muscle of clones versus controls at day 60 and day 260. We have analyzed the contractile and metabolic properties of muscle fibers by measuring the abundance of myosin heavy chain (MyHC) isoforms and activities of metabolic enzymes (LDH, PFK, COX, CS, ICDH), respectively. The spatial repartition of some components of the extracellular matrix (collagen types I, IV, VI, chondroitin-6-sulfate, decorin, and tenascin-X) was also studied. At day 60 we found lower numbers and structural organization of fibers, and a delay in the setup of the extracellular matrix. IGF-2 transcript abundance was lower in clones than in their controls. There was no difference in the expression of VEGF (a growth factor regulating vascularization and myogenesis) and its receptor. At day 260 the muscles of fetal clones have not reached the same degree of differentiation than controls as shown by their lower energy metabolisms and their MyHC pattern. These results show for the first time that disturbances in myogenesis occur early in fetal life in cloned cattle.