Search

This work investigated the metabolic basis for the variability of carcass and i.m. adiposity in cattle. Our hypothesis was that the comparison of extreme breeds for adiposity might allow for the identification of some metabolic pathways determinant for carcass and i.m. adiposity. Thus, 23- to 28-mo-old steers of 3 breeds, 2 with high [Angus or Japanese Black x Angus (J. Black cross)] and 1 with low (Limousin) i.m. and carcass adiposity, were used to measure activities or mRNA levels, or both, of enzymes involved in de novo lipogenesis [acetyl-coA carboxylase, fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme], circulating triacylglycerol (TAG) uptake (lipoprotein lipase), and fatty acid esterification (glycerol-3-phosphate dehydrogenase), as well as the mRNA level of leptin, an adiposity-related factor. In a first study, enzyme activities were assayed in the s.c. adipose tissue (AT), the oxidative rectus abdominis, and the glycolytic semitendinosus muscles from steers finished for 6 mo. Compared with Angus or J. Black cross, Limousin steers had a 27% less (P = 0.003) rib fat thickhess, and 23 and 29% less (P [less than or equal to] 0.02) FAS and G6PDH activities in s.c. AT. In rectus abdominis and semitendinosus, the 75% less (P < 0.001) TAG content was concomitant with 50% less (P < 0.001) G6PDH activity. In a second study, enzyme activities plus mRNA levels were assayed in an oxido-glycolytic muscle, the longissimus thoracis (LT), in the i.m. AT dissected from LT, and in s.c. AT from the same Limousin steers and from Angus steers finished for 10 mo. Compared with Angus, the 50% less (P < 0.001) rib fat thickness in Limousin contrasted with the 1.1- to 5.8-fold greater (P [less than or equal to] 0.02) mRNA levels or activities, or both, of acetyl-coA carboxylase, G6PDH, lipoprotein lipase, and glycerol-3-phosphate dehydrogenase in s.c. AT. Conversely, the 90% less (P < 0.001) TAG content in Limousin LT was concomitant to the 79 and 83% less (P [less than or equal to] 0.002) G6PDH activity and leptin mRNA level. Such differences could arise from a greater number of adipocytes in LT from Angus steers because no difference was found between Limousin and Angus for G6PDH activity and leptin mRNA in i.m. AT. We conclude that FAS and G6PDH in s.c. AT could be involved in differences in carcass adiposity, but this relationship disappeared when the fatness increased strongly. Leptin and G6PDH are related to the expression of marbling whatever the body condition and thus could be relevant indicators of marbling in beef cattle. Key words: adipose tissue, bovine, leptin, lipogenic activity, muscle

Marbling is an important criterion for beef quality grading in many countries. The purpose of the current study was to utilize the natural genetic variation to identify major metabolic indicators of marbling in cattle differing in genotypes. Rectus abdominis (RA, oxidative), semitendinosus (glycolytic), and longissimus thoracis (LT, oxido-glycolytic) muscles were taken from steers of different genotypes that expressed high [Angus, n = 16; and crossbred (Angus x Japanese Black), n = 10] or low (Limousin, n = 12) levels of marbling in their meat. Muscles from Angus and crossbred steers were characterized, as expected, by a greater triacylglycerol (TAG) content (P < 0.001) and also by greater protein contents of fatty acid-binding protein specific for heart and muscles (H-FABP; P < 0.001 for RA and P < 0.05 for LT muscle) or for adipocytes (A-FABP; P < 0.001 for RA and LT muscles). Moreover, oxidative enzyme activities ([beta]-hydroxyacyl-CoA dehydrogenase, citrate synthase, isocitrate dehydrogenase, cytochrome-c oxidase) were greater (P < 0.01 to 0.001) in the 3 muscles studied, whereas glycolytic enzyme activities (phosphofructokinase and lactate dehydrogenase) were lower (P < 0.001) in RA muscle in Angus and crossbred steers compared with Limousin steers. Significant correlations were observed between TAG content and H- and A-FABP protein contents, and oxidative (r [less than or eqaul to] +0.55, P < 0.001) or glycolytic enzyme activities (r [less than or equal] -0.47, P < 0.001), when the 3 genotypes and muscles studied were considered as a whole. In addition, A-FABP protein content and some oxidative enzyme activities were significantly correlated with TAG content independently of the genotype and muscle effects. In conclusion, A-FABP protein content, as well as oxidative enzyme activities, may be used as indicators of the ability of steers from extreme genotypes to deposit intramuscular fat. Key words: triacylglycerol, marbling, fatty acid-binding protein, muscle metabolism, genotype, steer

An experiment was conducted with 42 growing Montbeliard steers to study the effect of feed restriction, followed by refeeding, on circulating concentrations of thyroxine ([T.sub.4]) and triiodothyronine ([T.sub.3]) and on hepatic and muscle activities of 5'-deiodinase (5'D). At 9 mo of age, 21 steers were diet-restricted for 3 mo (ADG, 641 g/d), prior to a 4-mo compensatory growth period with ad libitum access to the same diet (ADG, 1,240 g/d). They were compared to 21 control steers continuously gaining 1,100 g/d between 9 and 16 mo of age. Blood samples were collected every 14 d and samples of liver and semitendinosus and triceps brachii (triceps) muscles were obtained at slaughter at the end of the restriction and refeeding periods (12 and 16 mo of age, respectively). Compared to control steers, feed restriction decreased plasma concentrations of ([T.sub.4]) after 56 to 83 d of feed restriction (P < 0.05), whereas ([T.sub.3]) concentration decreased only after 83 d of feed restriction (P < 0.05). No differences in hepatic and muscle 5'D activities were observed after 87 d of feed restriction and decreased growth rate (12 mo of age). During the refeeding period (compensatory growth), circulating concentrations of ([T.sub.4]) and ([T.sub.3]) were restored to control levels within 14 d. Moreover, ([T.sub.3]) concentration rose above that of control steers after 56 d of refeeding and remained higher for the duration of the experiment (P < 0.05). Hepatic 5'D activity was higher (P = 0.07) in compensated than in control steers at the end of refeeding period (16 mo of age) and higher (P < 0.01) after compensation at 16 mo than during restriction at 12 mo. Activities of 5'D in semitendinosus and triceps muscles were higher (P < 0.001) in 16-mo-old than in 12-mo-old steers, but no differences were observed due to feed restriction or compensatory growth. These results indicate that nutritional status regulates both thyroidal secretion and extrathyroidal ([T.sub.3]) production in cattle. The data also suggest that extrathyroidal ([T.sub.3]) production may be involved in the mechanism of compensatory growth in cattle. Key Words: Cattle, Compensatory Growth, Deiodination, Restricted Feeding, Thyroxine, Triiodothyronine

Myostatin, a member of the transforming growth factor-β superfamily, is a potent negative regulator of skeletal muscle growth and is conserved in many species, from rodents to humans. Myostatin inactivation can induce skeletal muscle hypertrophy, while its overexpression or systemic administration causes muscle atrophy. As it represents a potential target for stimulating muscle growth and/or preventing muscle wasting, myostatin regulation and functions in the control of muscle mass have been extensively studied. A wealth of data strongly suggests that alterations in skeletal muscle mass are associated with dysregulation in myostatin expression. Moreover, myostatin plays a central role in integrating/mediating anabolic and catabolic responses. Myostatin negatively regulates the activity of the Akt pathway, which promotes protein synthesis, and increases the activity of the ubiquitin-proteasome system to induce atrophy. Several new studies have brought new information on how myostatin may affect both ribosomal biogenesis and translation efficiency of specific mRNA subclasses. In addition, although myostatin has been identified as a modulator of the major catabolic pathways, including the ubiquitin-proteasome and the autophagy-lysosome systems, the underlying mechanisms are only partially understood. The goal of this review is to highlight outstanding questions about myostatin-mediated regulation of the anabolic and catabolic signaling pathways in skeletal muscle. Particular emphasis has been placed on (1) the cross-regulation between myostatin, the growth-promoting pathways and the proteolytic systems; (2) how myostatin inhibition leads to muscle hypertrophy; and (3) the regulation of translation by myostatin. [ABSTRACT FROM AUTHOR]

This paper reports the metabolic and morphological characteristics of bovine adipose tissue (AT) at end of fetal life and its variability with breed and anatomical site of AT. Our hypothesis was that, in cattle, end-of-fetal-life differences in adipocyte number, size, and histology may account for differences in AT maturity. To address this question, perirenal and intermuscular AT were sampled from Charolais, Blond d'Aquitaine, and Holstein fetuses at 260 d postconception. Holstein fetuses showed greater leptin mRNA abundance, which is consistent with the greater penrenal AT weight (P = 0.03) than Blond d'Aquitaine fetuses. Compared with Blond cl'Aquitaine or Charolais fetuses, Holstein fetuses had larger (P < 0.001) adipocytes, greater (P < 0.05) activities of enzymes involved in de novo fatty acid (FA) synthesis (FA synthase, glucose-6-phosphate dehydrogenase, malic enzyme) and FA esterification (glycerol-3-phosphate dehydrogenase), and greater (P = 0.06, P = 0.10, P < 0.001) mRNA abundance for lipolytic enzymes (hormone-sensitive lipase and adipose triglyceride lipase) and uncoupling protein 1 in both perirenal and intermuscular AT. This indicates increased FA turnover in Holstein aclipocytes through FA storage, mobilization, and oxidation pathways. Whatever the breed, adipocytes were smaller in penirenal AT than intermuscular AT. Whatever the breed or anatomical site, bovine AT at 260 d postconception contained predominantly unilocular adipocytes believed to be white adipocytes together with a few multilocular brown adipocytes. We conclude that the greater metabolic and morphologic maturity of adipocytes from Holstein than Blond d'Aquitaine and Charolais fetuses may contribute to the greater thermogeriic aptitude of Holstein newborns. 1 moreover, the presence of both white and brown adipocytes at the end of fetal life highlights the complexity of AT structure and may indicate that the cellular and functional heterogeneity of AT repeatedly observed postnatally has a developmental origin. [ABSTRACT FROM AUTHOR]

Myostatin (MSTN), a member of the TGF-β superfamily, is a negative regulator of skeletal muscle mass. We have previously shown that the cell survival/apoptosis pathway is a downstream target of MSTN loss-of-function in mice through the regulation of the expression or abundance of many survival and apoptotic factors. In this study, we used western-blot and quantitative PCR (qPCR) analyses to validate these novel downstream targets of MSTN in double-muscled (DM) cattle v. their controls including 260-day-old foetuses and adult cows from the INRA95 strain. MSTN loss-of-function in DM foetuses and DM cows resulted in a glycolytic shift of the muscles (e.g. upregulation of H-MyBP, PGM1 and SNTA1 and downregulation of H-FABP), activation of cell survival pathway through regulation of some components of the PI3K/Akt pathway (e.g. upregulation of DJ-1 and Gsk-3βser9/Gsk-3βtotal ratio and downregulation of PTEN) and upregulation of cell survival factors translationally controlled tumour protein (14-3-3E, Pink1). We also found a lower abundance of pro-apoptotic transcripts and/or proteins (Caspase-3, caspase-8, caspase-9, BID, ID2 and Daxx) and a higher expression of anti-apoptotic transcripts (Traf2 and Bcl2l2) in DM muscles. All together, these results are in favour of activation of the cell survival pathway and loss of apoptosis pathway within the muscles of DM animals. Alteration of both pathways may increase myonuclear or satellite cell survival, which is crucial for protein synthesis. This could contribute to muscle hypertrophy in DM foetuses and DM cows. [ABSTRACT FROM AUTHOR]

Abstract: The temporal relationships between thyroid status and differentiation of liver, heart and different skeletal muscles were examined in 42 bovine fetuses from day 110 to day 260 of development using principal component analysis of the data. Plasma concentrations of reverse-triiodothyronine (rT3) and thyroxine (T4) increased during development from day 110 to day 210 or 260, respectively, whereas concentration of triiodothyronine (T3) and hepatic type-1 5′-deiodinase activity (5′D1) increased from day 180 onwards. On day 260, high T4 and rT3 and low T3 concentrations were observed together with a mature 5′D1 activity. Cytochrome-c oxidase (COX) activity expressed per mg protein increased at day 180 in masseter and near birth in masseter, rectus abdominis and cutaneus trunci muscles (P <0.05). Significant changes in citrate synthase (CS) activity per mg protein were observed between day 110 and day 180 in the liver and between day 210 and day 260 in the liver, the heart and the longissimus thoracis muscle (P <0.05). Muscle contractile differentiation was shown by the disappearance of the fetal myosin heavy chain from day 180 onwards. A positive correlation (r >0.47, P <0.01) was shown between thyroid status parameters (5′D1, concentrations of T4 and T3) and COX activity in muscles known to be oxidative after birth (masseter, rectus abdominis) but not in liver and heart, nor in muscles known to be glycolytic after birth (cutaneus trunci, longissimus thoracis). A similar correlation was found between thyroid parameters and CS activity in liver and masseter. Results indicate that elevation of plasma T3 concentrations in the last gestational trimester could be involved in the differentiation of oxidative skeletal muscles. [Copyright &y& Elsevier]

An experiment was conducted with 42 Montbéliard steers to determine if nutritionally induced interrupted growth could influence muscle characteristics of steers and hence meat quality. A restriction/refeeding path was designed in order to induce a discontinuous growth path. At 9 months of age, 21 steers were given a restricted amount of diet for 3 months and were then slaughtered (R steers; no. = 10; intake: 5.28 kg dry matter (DM) per day) or sgerowthubjected to a 4-month ad libitum refeeding period (R/F steers; no. = 11; intake: 8.99 kg DM per day) with the same diet (11.03 to 11.12 MJ metabolizable energy (ME) per kg DM) prior to slaughter. An additional 21 control steers were offered the same diet but in amounts that allowed them to gain continuously between 9 and 12 months of age, and were then slaughtered (C steers; no. = 10; intake: 7.08 kg DM per day) or maintained on a continuous feeding protocol through to 16 months of age prior to slaughter (C/C steers; no. = 11; intake : 8.07 kg DM per day). M. semitendinosus (ST), m. longisssimus thoracis and m. triceps brachii (TB) were collected for biochemical and histochemical analyses. R steers had a lower average daily gain (ADG; P < 0.001), a lower final weight (P < 0.01) and a leaner carcass (P < 0.01) than C steers. Upon refeeding, R/F steers had a higher ADG than C/C steers (P < 0.05) and underwent compensatory growth resulting in compensation of body weight and composition at 16 months. In muscles, glycolytic lactate dehydrogenase activity was lower in R steers (P < 0.01) and restored in R/F steers compared with control steers. Among oxidative enzymes, cytochrome-c oxidase activity was higher in the TB of R/F compared with C/C steers (P < 0.001) indicating a muscle-specific metabolic adaptation to the feeding level. There was little effect of the nutritional treatment on muscle fibre size and type except for an increase in the frequency of hybrid fibres in R and R/F groups... [ABSTRACT FROM AUTHOR]

Evaluates the effect of food restriction and refeeding associated with reduced and compensatory growth on circulating concentrations of thyroid hormones and activity of type-I 5'-deidinase in liver and muscle of steers. Evolution of circulating concentrations of thyroid hormones in preruminant, weaned and growing steers; Influence of weaning on circulating concentrations of thyroid hormones; Influence of feeding level in the postweaning growth period.

Information on clone offspring F1 is limited, especially in species with a long inter-generation interval such as cattle. As cloned cattle exhibit a slight delay in muscle maturation until puberty (Jurie et al.2009 Animal 3, 244-250), the present study aimed to investigate the contractile and metabolic muscle characteristics of F1 at 8, 12, and 18 months of age. Repeated biopsies of the semitendinosus muscle were collected on 10 F1 heifers born after AI of cloned cows at the experimental farm of INRA. Muscle characteristics of these offspring were compared with those of 9 female clones and 8 AI control heifers previously biopsied at the same ages. All animals (clones, F1, and controls) were female Holstein, born and raised under the same conditions in the same farm. Biopsy samples were stored frozen at -80°C until analysis for contractile and metabolic characteristics. The type of contractile fibers was determined from the proportion of the different myosin heavy chain (MyHC) isoforms separated by electrophoresis. Oxidative metabolism was assessed by isocitrate dehydrogenase (ICDH) and cytochrome-c oxidase (COX) activities (μmol min-1per gram of muscle). Data were analyzed separately for each time of biopsy using the GLM procedure of SAS (SAS Institute, Cary, NC, USA). The statistical model contained the group as fixed effect. When a significant effect was detected, differences between least squares means were further separated by the PDIFF option of SAS. Comparison of contractile characteristics from the 3 groups of animals is presented in Table 1. The proportion of MyHC I (slow oxidative isoform) and MyHC IIx (fast glycolytic isoform) in the muscles of F1 was not significantly different from those of controls at 8 and 12 months of age. F1 had different muscle contractile properties compared with clones at 12 months of age. At 8 months of age, F1 had greater ICDH activity than controls (1.39 ± 0.22 v.0.54 ± 0.007; P≤ 0.002) and greater COX activities (11.4 ± 1.6 v.4.2 ± 0.9; P≤ 0.003), but this was not observed later on. Altogether, these data indicate that the muscles of F1 were more oxidative than those of controls. This was not related to a higher percentage of type I fibers but rather to a higher percentage of type IIA fibers. A delay in muscle maturation was only partially found in F1. Table 1.Contractile characteristics (mean ± SE) of the semitendinosus muscle from controls (n= 8), clones (n= 9) and clone offspring (F1, n= 10) heifers [ABSTRACT FROM AUTHOR]

Previous data have shown that bovine clones display a delay in their muscle differentiation during their first year postnatal (Jurie et al.2009 Animal 2, 244-250). This delay could originate from perturbations in fetal muscle development as illustrated by lower numbers and degree of organization of the first generation of myotubes at 60 dpc and by their lower energy metabolism and their myosin heavy chain pattern at 260 dpc (Cassar-Malek et al.2009 Proc. XIth ISRP abst). In order to understand the mechanisms underlying the delay in myogenesis, we have performed a comparative proteomic analysis of the semitendinosus muscle in fetuses derived from somatic nuclear transfer and their control counterparts obtained after AI at these two important developmental stages. Two-dimensional electrophoresis using a 3-10 non-linear pH gradient were performed on samples at 60 dpc (in a group of Holstein animals and a group of Charolais animals, n= 4 fetuses per lot) and at 260 dpc (in a group of Holstein animals, n= 4 fetuses per lot). Gel analysis was conducted using the image analysis SameSpots (Nonlinear Dynamics, Newcastle upon Tyne, UK). As expected, the protein profiles were visually very different between developmental stages. At 60 dpc, 463 spots common to all gels were retained for statistical analysis using the significance analysis of microarrays (SAM) method (FDR <5%; Meunier et al.2005 Anal. Biochem. 340, 226-230). At 260 dpc, 491 spots were selected for SAM analysis. The statistical analysis revealed a small number of differential spots (9 and 10 spots, respectively, at 60 dpc in Holstein and Charolais, and 10 spots at 260 dpc Holstein). The differential spots were excised from the gels and their identification by mass spectrometry is in progress. Preliminary results are presented in Table 1. In conclusion, subtle changes in the muscle proteome were detected in clones v.controls. Some of them were related to the regulation of cell cycle/apoptosis at 60 dpc and to energy metabolism and chaperone activity at 260 dpc. The relevance of these changes will be further explored using bioinformatics tools. Table 1.Examples of identified spots with differential abundance between clones and controlsThe authors thank C. Barboiron for excellent technical assistance. [ABSTRACT FROM AUTHOR]

The knowledge of the molecular organization of living organisms evolved considerably during the last years. The methodologies associated also progressed with the development of the high-throughput sequencing (SNP array, RNAseq, etc.) and of genomic tools allowing the simultaneous analysis of hundreds or thousands of genes, proteins or metabolites. In farm animals, some proteins, mRNAs or metabolites whose abundance has been associated with meat quality traits have been detected in pig, cattle, chicken. They constitute biomarkers for the assessment and prediction of qualities of interest in each species, with potential biomarkers across species. The ongoing development of rapid methods will allow their use for decision-making and management tools in slaughterhouses, to better allocate carcasses or cuts to the appropriate markets. Besides, their application on living animals will help to improve genetic selection and to adapt a breeding system to fulfill expected quality level. The ultimate goal is to propose effective molecular tools for the management of product quality in meat production chains. [ABSTRACT FROM AUTHOR]

Abstract: Array technology has been increasingly used to monitor global gene expression patterns in various tissues and cell types. However, applications to muscle development and pathology as well as meat production in livestock species have been hampered by the lack of appropriate cDNA collections. To overcome this problem, a directed cDNA library was constructed starting from 23 muscles of meat-producing bovines to derive a collection of 3573 clones. A preliminary sequence characterization of this collection indicated that the most abundant transcripts correspond to genes encoding proteins involved in energy metabolism (COX and NADH dehydrogenase subunits) and belonging to the contractile apparatus (myosin chains and troponin isoforms). From this cDNA library, we selected a set of 435 clones representing 340 unique genes, of which 24 were novel. This collection was subsequently completed with 75 specific cDNA probes for genes of interest already studied in our laboratory. The bovine ‘muscle’ cDNA repertoire thus designed was spotted onto a nylon membrane (macroarray) in order to test its utility to further investigate the transcriptome of bovine muscles in relation to meat quality traits. It is also anticipated that this type of collection might be useful for the study of chronic myologic diseases in other mammalian species, including humans. [Copyright &y& Elsevier]