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Intrauterine growth restriction (IUGR) occurs naturally in pigs and leads to low birth weight of piglets due to undernutrition caused by placental insufficiency. For 2 main reasons, low birth weight causes economic loss. First, low birth weight pigs have a greater mortality and increasing the litter size causes more low birth weight piglets within litters. Second, surviving low birth weight piglets have reduced performance (i.e., ADG, feed conversion rate, and percentage meat). To develop dietary strategies for preventing IUGR, knowledge of the biological basis of IUGR is required. Muscle fiber number, formed during myogenesis, is correlated positively with performance traits and has been shown in several studies to be reduced in low birth weight pigs. Postnatal muscle hypertrophy is due to satellite cell number per fiber at birth and their rate of proliferation as well as protein deposition (i.e., protein synthesis and degradation). Previous studies and some recent ones indicate that low birth weight littermates in mice are born with fewer satellite cells and studies on pigs show that the rate of satellite cell proliferation may vary within litters. Proteomics studies show that protein synthesis and degradation is downregulated in IUGR pigs and low birth weight pigs also produce meat with less tenderness. Alternative maternal feeding strategies to prevent IUGR have been examined. Increasing maternal global nutrition had no beneficial effect on performance and muscle growth traits in several studies. Feeding excess maternal dietary protein also did not influence muscle growth traits whereas moderately decreased maternal dietary protein may decrease muscle fiber number and performance. On the other hand, addition of L-carnitine to the maternal gestation or lactation diet may increase birth and weaning weights or the muscle fiber number, respectively, in low birth weight pig offspring. Finally, promising data have been obtained on reproductive traits in pigs after addition of functional AA, such as arginine and glutamine, to the gestational diet. Although much is known about the biological basis of IUGR, we still need to learn much more about the mode of action before maternal dietary strategies can be developed to prevent IUGR.

Acute physical activity elicits changes in gene expression in skeletal muscles to promote metabolic changes and to repair exercise-induced muscle injuries. In the present time-course study, pigs were submitted to an acute bout of treadmill running until near exhaustion to determine the impact of unaccustomed exercise on global transcriptional profiles in porcine skeletal muscles. Using a combined microarray and candidate gene approach, we identified a suite of genes that are differentially expressed in muscles during postexercise recovery. Several members of the heat shock protein family and proteins associated with proteolytic events, such as the muscle-specific E3 ubiquitin ligase atrogin-1, were significantly upregulated, suggesting that protein breakdown, prevention of protein aggregation and stabilization of unfolded proteins are important processes for restoration of cellular homeostasis. We also detected an upregulation of genes that are associated with muscle cell proliferation and differentiation, including MUSTN1, ASB5 and CSRP3, possibly reflecting activation, differentiation and fusion of satellite cells to facilitate repair of muscle damage. In addition, exercise increased expression of the orphan nuclear hormone receptor NR4A3, which regulates metabolic functions associated with lipid, carbohydrate and energy homeostasis. Finally, we observed an unanticipated induction of the long non-coding RNA transcript NEAT1, which has been implicated in RNA processing and nuclear retention of adenosine-to-inosine edited mRNAs in the ribonucleoprotein bodies called paraspeckles. These findings expand the complexity of pathways affected by acute contractile activity of skeletal muscle, contributing to a better understanding of the molecular processes that occur in muscle tissue in the recovery phase.

In the present study, the metabolic effects of heat and anoxic stress in myotubes from the mouse cell line C2C12 were investigated by using a combination of (13)C, (1)H, and (31)P nuclear magnetic resonance (NMR) spectroscopy and enrichment with [(13)C]-glucose. Both the (13)C and the (1)H NMR spectra showed reduced levels of the amino acids alanine, glutamate, and aspartate after heat or anoxic stress. The decreases were smallest at 42 degrees C, larger at 45 degrees C, and most pronounced after anoxic conditions. In addition, in both the (1)H and the (31)P NMR spectra, decreases in the high-energy phosphate compounds adenosine triphosphate and phosphocreatine with increasing severity of stress were identified. At anoxic conditions, an increase in (13)C-labeled lactate and appearance of glycerol-3-phosphate were observed. Accumulation of lactate and glycerol-3-phosphate is in agreement with a shift to anaerobic metabolism due to inhibition of the aerobic pathway in the mitochondria. Conversely, lower levels of unlabeled ((12)C) lactate were apparent at increasing severity of stress, which indicate that lactate is released from the myotubes to the medium. In conclusion, the metabolites identified in the present study may be useful markers for identifying severity of stress in muscles., (Copyright 2010 Elsevier Inc. All rights reserved.)

The aim of this study was to investigate the effect of pre-slaughter fasting time, sex and feeding regime on the development of energy metabolism and pH in M. longissimusdorsi (LD) post-mortem in pigs. Two hundred and seventy pigs of the commercial Norwegian crossbreed Noroc (LYLD) were used involving two sexes (gilts and castrates), two feeding regimes (restricted and ad libitum) and four different fasting treatments: (F4) 4h fasting, (F175) 17.5h fasting on the farm, (FO175) 17.5h fasting overnight at the abattoir, and (FO265) 26.5h fasting overnight at the abattoir. Additionally the pigs experienced two different abattoir lairage times as fasting treatment F4 and F175 had a lairage time of 1.5h, while fasting treatment FO175 and FO265 had a lairage time of 23.0 h. A short fasting time of 4 h led to a delayed degradation of glycogen, slow decline in pH and a lower ultimate pH(45 h) post-mortem (pHu) in the LD compared with a fasting time of 26.5 h which resulted in a rapid breakdown of glycogen and pH decline early post-mortem and a high pHu. Proglycogen was degraded in favour of macroglycogen under anaerobic conditions post-mortem. Feeding the animals in the morning before delivery if slaughtered the same day, results in low pH reduction rate and a low pHu compared with pigs fasted overnight either on farm or at the abattoir. Aiming a higher pHu in LD it should be recommended not to feed the pigs in the morning at the day of slaughter.

To establish an adequate model to study the proliferation and differentiation of porcine skeletal muscle in response to bioactive compounds, a pool of satellite cells was derived from the semimembranosus muscle (SM) of newborn piglets using a Percoll gradient centrifugation. The final yield amounted to 4.1 x 10(6) cells/g muscle tissue. The percentage of muscle satellite cells has been determined by immunostaining for desmin and subsequent fluorescence analysis by flow cytometry, which revealed 95% of desmin-positive cells. For proliferation studies, satellite cell born myoblasts were seeded in gelatin-coated 96-well microplates at about 5 x 10(3) cells per well. Cells were grown for 1 day in MEMalpha plus 10% fetal bovine serum (FBS) and 10% horse serum (HS), followed by 2 d cultivation in serum-free growth medium. For differentiation studies, myoblasts were cultured in matrigel-coated 24-well plates for 4 d with growth medium containing 10% FBS and 10% HS. At 80% confluence, cells were grown for 24 h in medium plus 10% FBS and 1 microM insulin to initiate differentiation. Subsequently, the cells were cultured in serum-free differentiation medium (SFDM) for 3 d to form myotubes. Cultures reached a maximum fusion rate of approximately 20% after 96 h. By establishing this culture system, we provide an advanced and appropriate in vitro model to study porcine skeletal muscle cell growth and differentiation including the responses to various bioactive compounds.

The role of phospholipase A2 in the induction of drip loss from pig muscle has been investigated. In samples from porcine M. longissimus dorsi, total PLA2 activity as well as mRNA and protein levels of the group VIA iPLA2 (iPLA2-VIA) increased during the initial 4 h post-mortem period. Morphological studies of porcine muscle showed that at 4 h post-mortem, gaps had formed between muscle fibers and that the sarcolemma membrane borders appeared blurred. At the same time iPLA2-VIA protein levels were increased inside muscle fibers and at the sarcolemma. iPLA2-VIA mRNA abundance in samples from different breeds of pigs with variations in drip loss revealed no clear correlation between drip loss level and iPLA2-VIA expression. Together, these data indicate that during the post-mortem period, iPLA2-VIA expression and activity is increased at the muscle fiber membranes. PLA2 activity may affect membrane permeability and consequently the progression of drip formation in porcine muscle.

Sixteen female pigs (Duroc x Landrace x Large White) were divided into 2 groups, which had either free access to the diet (control group) or were feed-restricted from d 28 to 80 and then had free access to the diet (compensatory growth group). The sensory analysis showed that the pigs exhibiting compensatory growth produced meat with increased tenderness compared with control pigs (P < 0.05). To gain further knowledge of the influence of compensatory growth on meat tenderness, the sarcoplasmic protein fraction of muscle tissue was studied at the time of slaughter and 48 h postmortem using proteome analysis. At slaughter, 7 different proteins were found to be affected by compensatory growth: HSC70, HSP27, enolase 3, glycerol-3-phosphate dehydrogenase, aldehyde dehydrogenase E2, aldehyde dehydrogenase E3, and biphosphoglycerate mutase. The HSC70 and HSP27 both belong to the heat shock family and are known to play a role during muscle development. Hence, they may be affected by compensatory growth and increased protein turnover. Forty-eight hours after slaughter, 8 different proteins were found to be affected by compensatory growth: myosin light chain (MLC) II, MLC III, sulfite oxidase, chloride intracellular channel 1, 14-3-3 protein gamma, elongin B, and phosphohistidine phosphatase 14. The changes observed on MLC II and MLC III could be a consequence of enzymatic cleavage in the neck region of the globular myosin head domain that causes the release of MLC II and MLC III from the actomyosin complex. It has previously been hypothesized that compensatory growth results in an increased postmortem proteolysis; thus it was presumed that the intensity of some protein fragments would be affected by compensatory growth. However, the peptides that were found to be affected at 48 h postmortem were all full-length proteins. The 14-3-3 protein gamma has been proposed to play a role in the contraction of muscle during rigor and may thereby have an effect on meat tenderness. This study reveals some very interesting changes in the muscle proteome affected by compensatory growth, which may be useful in understanding the relationship among compensatory growth, protein turnover, and meat tenderness.

The objective of this study was to examine the intralitter variation in postnatal growth performance, meat quality, and muscle fiber characteristics when littermates were categorized by carcass weight. Thirty-nine litters were weaned at 4 wk of age and had free access to feed from 2 wk of age until slaughter. They were slaughtered by litter at an average BW of 104 +/- 14 kg, and six pigs per litter were selected for analysis: the heaviest- (HW), middle- (MW), and lightest-weight (LW) pig of each sex. Categorizing littermates in LW, MW, and HW pigs at the same age reflected the differences in postnatal growth rate within a litter; thus ADG, muscle mass, and muscle deposition rate differed across pig weight groups (P < 0.001). Also, the total DNA content was different among pig weight groups (P < 0.001) and reflected differences in muscle growth rate. The difference in muscle growth rate between LW and MW pigs could be explained by a larger (P < 0.05) mean fiber area (MFA) in MW pigs, whereas the number of muscle fibers was similar. Growth rate differences between MW and HW pigs could in part be explained by a higher number (P < 0.01) of equal-sized muscle fibers in HW pigs. The difference in MFA was due to a higher estimated DNA and RNA content per muscle fiber in MW and HW compared with LW pigs (P < 0.05). Pigment content was higher in MW and HW compared with LW pigs (P < 0.01), but no other measured meat quality traits were significantly different across pig weight groups. These results indicate that both the number and the growth rate of muscle fibers contribute to intralitter variation in postnatal growth performance.

The objective of this study was to examine how increased feed intake of the sow during early to mid-gestation affects sow performance and the muscle fiber number, performance, and technological meat quality of the offspring. Thirty-nine pregnant sows (Landrace x Large White sows mated to Landrace or Large White boars) in their fourth parity were assigned to one of three treatments: 1) the sows were either fed restrictively (control = 15 MJ of NE/d from d 1 to 90, then 24 MJ of NE/d from d 91 to 112, and again 15 MJ of NE/d from d 113 to 115 of gestation); 2) fed ad libitum from d 25 to 50 (A25-50); or 3) ad libitum from d 25 to 70 (A25-70) and as control in the remaining periods. The offspring were weaned at 4 wk of age and had free access to feed from 2 wk of age until slaughter. They were slaughtered litterwise at an average body weight of 104 +/- 14 kg. Estimates for total, primary (P-), and secondary (S-) muscle fiber number; muscle fiber area; and DNA and RNA content were analyzed in semitendinosus muscle (ST) samples from the heaviest, middle, and lightest weight (LW) pigs of each sex within litter selected at slaughter. Technological meat quality traits (pH at 24 h postmortem, drip loss, Minolta color, and pigment) were analyzed in longissimus dorsi muscle. Fiber number, fiber area, and concentrations and content of DNA and RNA of the offspring were not significantly affected by increased maternal nutrition. The ST muscle weight was lower in offspring from A25-50 than control sows (P = 0.019). Average daily gain, carcass weight, and the muscle deposition rate also were numerically lower for A25-50 than control and A25-70 pigs. An interaction between treatment and pig weight was found for muscle deposition rate (P = 0.006), in that LW pigs from treatment A25-50 had a lower deposition rate than LW pigs from control. We found no effect of treatment on the meat quality traits in the offspring. Also, barrows had a higher (P < 0.05) number of P-fibers, higher daily gain, and carcass weight than female pigs. No differences were found on any meat quality traits between sexes. Thus, ad libitum feeding of pregnant sows from d 25 to 50 or d 25 to 70 of gestation did not have any beneficial effect on muscle fiber number and area in the offspring. It seems that maternal ad libitum feeding from d 25 to 50 in gestation had a negative effect on postnatal muscle growth, with especially the LW pigs being affected.

The present study illustrates elements of the signal cascades involved in the activation of taurine efflux pathways in myotubes derived from skeletal muscle cells. Exposing primary skeletal muscle cells, loaded with (14)C-taurine, to 1) hypotonic media, 2) the phospholipase A(2) (PLA(2)) activator melittin, 3) anoxia, or 4) lysophosphatidyl choline (LPC) causes an increase in (14)C-taurine release and a concomitant production of reactive oxygen species (ROS). The antioxidants butulated hydroxy toluene and vitamin E inhibit the taurine efflux after cell swelling, anoxia, and addition of LPC. The muscle cells possess two separate taurine efflux pathways, i.e., a swelling- and melittin-induced pathway that requires 5-lipoxygenase activity for activation and a LPC-induced pathway. The two pathways are distinguished by their opposing sensitivity toward the anion channel blocker DIDS and cholesterol. These data provide evidence for PLA(2) products and ROS as key mediators of the signal cascade leading to taurine efflux in muscle.

The effects of various growth rates in pigs induced by four different feeding strategies on the activity of the calpain system and on postmortem (PM) muscle proteolysis and tenderness development were studied. An increased growth rate may be caused by an increased protein turnover, which results in up-regulated levels of proteolytic enzymes in vivo that, in turn, possibly will affect PM tenderness development. It can be hypothesized that increased proteolytic activity pre-slaughter will increase the PM tenderization rate. From postnatal d 28 to d 90 (phase 1) the pigs were divided into two groups, given either ad libitum (A) or restricted (R, 60% of ad libitum) access to feed. The two groups were then divided into two subgroups, given either restricted or ad libitum access to feed from d 91 to slaughter at d 165 (phase 2). Measurements of the activity of mu-calpain, m-calpain, and calpastatin; concentrations of total collagen and the percent of soluble collagen; and RNA, DNA, and elongation factor-2 where made at slaugther. Myofibrillar fragmentation index (MFI) was determined at slaughter and 24 h PM. Warner-Braztler shear force was determined 1 d and 4 d PM. Pigs fed restricted diets in phase 1 and fed ad libitum in phase 2 (RA pigs) had increased growth rates in the last phase compared to pigs fed ad libitum during both phase 1 and phase 2 (AA pigs). The increased growth rate (compensatory growth) was followed by an increased proteolytic potential (mu-calpain:calpastatin ratio), increased MFI values, and higher tenderization rates. There was a positive correlation between the activities of m-calpain and growth rates (r = 0.35, P = 0.03), and between RNA levels and growth rates (r = 0.43, P = 0.006). The proposed hypothesis is largely supported by the results. The activities of both mu- and m-calpain at slaughter were highest in fast-growing pigs. The calpain activity was highest in RA pigs, which in turn also had the fastest growth rates prior tslaughter among the four groups. This implies that the synthesis of these enzymes was up-regulated during the second feeding period to a larger extent in RA pigs. The proteolytic potential and the MFI values indicate that the up-regulated in vivo calpain activity had an effect on PM protein degradation, which also is supported by the higher tenderization rate in RA pigs.

This study investigates the effects of developmental stage and muscle type on the mobility and distribution of water within skeletal muscles, using low-field (1)H-NMR transverse relaxation measurements in vitro on four different porcine muscles (M. longissimus dorsi, M. semitendinosus, M. biceps femoris, M. vastus intermedius) from a total of 48 pigs slaughtered at various weight classes between 25 kg and 150 kg. Principal component analysis (PCA) revealed effects of both slaughter weight and muscle type on the transverse relaxation decay. Independent of developmental stage and muscle type, distributed exponential analysis of the NMR T(2) relaxation data imparted the existence of three distinct water populations, T(2b), T(21), and T(22), with relaxation times of approximately 1-10, 45-120, and 200-500 ms, respectively. The most profound change during muscle growth was a shift toward faster relaxation in the intermediate time constant, T(21). It decreased by approx. 24% in all four muscle types during the period from 25 to 150 kg live weight. Determination of dry matter, fat, and protein content in the muscles showed that the changes in relaxation time of the intermediate time constant, T(21), during growth should be ascribed mainly to a change in protein content, as the protein content explained 77% of the variation in the T(21) time constant. Partial least squares (PLS) regression revealed validated correlations in the region of 0.58 to 0.77 between NMR transverse relaxation data and muscle development for all the four muscle types, which indicates that NMR relaxation measurements may be used in the prediction of muscle developmental stage.

The objectivity of two of the most widely used methods for differentiation of fibre types, i.e. 1) the myosin ATP-ase method (Brooke and Kaiser, 1970a,b) and 2) the combined method, by which the myosin ATP-ase reaction is used to differentiate between fast and slow twitch fibres and NADH-tetrazolium reductase activity is used to identify the subgroups of fast twitch fibres (Ashmore and Doerr, 1970, Peter et al., 1972), was assessed in muscle samples from horses, calves and pigs. We also assessed the objectivity of the alpha-amylase-PAS preparation for the visualisation of capillaries (Andersen, 1975) in these species. For the purpose of reducing the time costs of histochemical analysis of muscle samples, we have developed an interactive image analysis system which is described. All analyses are performed on this system. In accordance with several other investigations, differences between the two methods of differentiating fibre types were found only for the relative distribution of the fast-twitch fibre subgroups (p < 0.02 and p < 0.05 for the relative number of type IIA and IIB, respectively) whereas when calculated in relative area, only the proportion of type IIA fibres differed significantly (p < 0.05). Significant interaction effects between method and person and method and species were found for the same traits. When methods were analysed separately, the combined method displayed a significant variance component of person for the distribution of type IIA fibres both in relative number and in relative area (p < 0.01 and p < 0.001, respectively). The objectivity of the ATP-ase method was further substantiated by the fact that a higher fraction of the total variance for the relative area distribution of type IIA fibres was explained by the statistical model (72.1% versus 34.2% for the combined method). Although the repeatability of measurements between persons was generally high (> 87%), the impact of differences in pre-requisites (varied degrees of overlap between the fibre types) for performing the differentiation by the combined method raises a question of the reliability of this method. Apparently, no general rules for comparison of results of distribution of the two subgroups of fast twitch fibres by the two methods are applicable. The alpha-amylase-PAS method was found to be a fairly objective method to identify capillaries in muscles from horses, calves and pigs. However, as capillarity described in combination with other traits to give an indication of diffusion characteristics is significantly influenced by person, it is recommended that the same person perform all the analysis of a project. In addition to the methodological results in this study, we have shown that by application of the TEMA image analysis system, which is more rapid compared with the time-consuming traditional method for evaluation of histochemical preparations, analysis of samples from large-scale experiments are now possible.

The objective of this study was to determine the effect of the beta-adrenergic agonist cimaterol (CIM) on fiber characteristics, capillary supply, and metabolic enzyme activities in muscles of young Friesian bulls. Four pairs of monozygotic twins in each of three live weight groups (WG) were used (initial average live weight [LW]: 162, 299, and 407 kg, respectively). Within each pair, one twin was fed .06 mg of CIM.kg LW-1.d-1 for 90 d. The other twin served as control (C). Needle biopsies were obtained from the longissimus (LM) and semitendinosus (ST) muscles at d 82 to 84 of treatment, and muscle fibers were identified as slow-twitch (Type I) or fast-twitch (Type IIA or Type IIB) by the myosin ATPase stain. In LM, the proportion of Type I (C: 24.0%, CIM: 20.4%; P < .07) and Type IIA fibers (C: 24.2%, CIM: 8.6%; P < .001) decreased, whereas the proportion of Type IIB fibers increased (C: 51.7%, CIM: 71.1%; P < .001). Cimaterol increased the cross-sectional area of Type I (P < .02) and Type IIB fibers (P < .001), with no change in Type IIA fibers. Overall, the mean fiber area increased (C: 2,363 microns 2, CIM: 3,934 microns 2; P < .001). The number of capillaries per fiber did not change, but the number of capillaries per square millimeter decreased (P < .001) after CIM treatment. Cimaterol changed metabolic enzyme activities toward lower oxidative capacity of the muscle (lactate dehydrogenase: +22%, hydroxyacyl-CoA dehydrogenase: -33%, and citrate synthetase: -34%; all P < .001) and reduced the glycogen content by 25% (P < .01).(ABSTRACT TRUNCATED AT 250 WORDS)