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1.
The activities of calpain and caspase systems during ageing in Longissimus lumborum (LL) and Infraspinatus (IS) muscles of Italian Simmental young bulls (Bos taurus) were assessed. Samples from 10 animals were collected within 20 min of exsanguination (T0), after 48 h (T1) and 7 days (T2) post mortem. Calpain and caspase activity were evaluated based on the formation of αII spectrin cleavage products of 145 kDa (SBDP145) and 120 kDa (SBDP120), respectively. Caspase activity was also assessed by the presence of poly (adenosine diphosphate‐ribose) polymerase‐1 (PARP‐1) cleavage product. At T0, LL showed higher levels of SBDP145 than IS (P < 0.01), while SBDP120 and PARP‐1 degradation products were similar between muscles. At T1, no difference was found in the level of SBDP145 between muscles, while SBDP120 and PARP‐1 cleavage products were not detected. At T2 neither αII spectrin nor PARP‐1 cleavage products were found. LL and IS showed different proteolysis after slaughter that was influenced more by calpain than caspase activity, which was detectable only in the early post mortem period.  相似文献   

2.
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3.
The objective of this study was to examine the differences in calpain system, desmin degradation, pH values and water holding capacity (WHC) between muscles of commercial Meishan and Duroc × Landrace × Yorkshire crossbred pigs. Meishan pork presented better WHC evidenced by lower purge loss at days 1 and 3 and less centrifugation loss at day 1 post mortem (P < 0.05). pH values at 45 min post mortem in Meishan pork were significantly higher compared to crossbred pork (P < 0.05). Calpain‐1 messenger RNA (mRNA) expression was lower in Meishan pork compared to that from crossbred pork (P < 0.05). Additionally, calpain‐1 activity, the ratio of calpain‐1 to calpastatin activity and desmin degradation were lower in Meishan pork compared to those from crossbred pork samples (P < 0.05). The results indicate that the calpain system including mRNA expression and activity were different between commercial Meishan and crossbred pork resulting in difference in the degree of desmin degradation during post mortem aging. pH values at 45 min and 24 h post mortem rather than calpain activity and desmin degradation could explain the higher water holding capacity in commercial Meishan pork.  相似文献   

4.
The biochemistry of intermuscular variation in tenderness is not fully understood. To investigate the role of the calpains in this process we performed two experiments using bovine and ovine species. In the bovine experiment, two distinct muscles, longissimus thoracis et lumborum (LT) and psoas major (PM), were used. In the ovine experiment, four muscles, LT, PM, semimembranosus (SM), and semitendinosus (ST), were used. Muscles were sampled at death for the determination of the steady-state mRNA level of calpains and calpastatin and the activities of calpain 1, 2, and calpastatin. Muscles were also sampled to determine the temporal changes in pH, tenderness, and the activity of the ubiquitous calpain system during postmortem aging. The results of the relative rate of tenderization in both species was found to be related to muscle type; LT had the highest value in both species. Within species, the mRNA steady-state levels of calpain 1 and calpastatin were similar in various bovine and ovine muscles. Bovine calpain 2 mRNA level was significantly lower in the LT than in the PM. Ovine calpain 2 mRNA level was lower, but not significantly different, in the LT compared to the other muscles. The mRNA level of bovine calpain 3 was significantly higher in the LT muscle than in the PM. In the ovine, the mRNA level of calpain 3 was highest in the LT, followed by SM, PM, and ST. Results on the activity of the ubiquitous calpain system in various muscles at death were dependent on muscle type and species. Temporal changes in the activity of calpains and calpastatin during the first 24 h of postmortem aging were similar in the muscles studied: calpain 1 and calpastatin declined significantly and calpain 2 remained relatively unchanged. The temporal changes in muscle pH in both experiments indicated that the extent and rate of pH decline during aging was related to muscle type. Correlation analysis between the relative rate of tenderization and mRNA expression of calpains revealed a strong relationship with calpain 3 in both species.  相似文献   

5.
The present study was conducted to elucidate the effect of dietary lysine levels on the intramuscular fat (IMF) content in the Longissimus dorsi (L. dorsi) muscles of finishing gilts. Eleven gilts in total from two litters of pigs aged 110 days were used. The average initial bodyweight of the pigs was 61.7 kg. Six pigs were assigned to the low lysine (LL) diet group (lysine content: 0.43 or 0.40%) and five pigs were assigned to the control group (lysine content: 0.65 or 0.68%). The diets were iso‐energetic and iso‐protein, and contained all essential amino acids (apart from lysine) in the recommended amounts. The pigs were fed these diets until their live weights reached 110 kg. Live weight gain and feed efficiency tended to be lower in the LL group (P = 0.118 and P = 0.052, respectively). Pigs from the LL group took 5 days longer to reach 110 kg (P < 0.01). The IMF content in the L. dorsi of the LL group was twice as high as that of the control group (6.7 vs 3.5%; P < 0.01). The percentage of oleic acid in the L. dorsi of the LL group tended to be higher than that of the control group (P = 0.052), whereas the percentage of linoleic acid and the total percentage of polyunsaturated fatty acids in the L. dorsi were lower (P < 0.05) in the LL group. Free L‐carnitine content in the L. dorsi was lower (P < 0.05) in the LL group. The average abundance of peroxisome proliferator‐activated receptor gamma mRNA in the L. dorsi of the LL group was threefold higher than that of the control group. The leptin mRNA abundance in the L. dorsi of the LL group was 3.3‐fold higher than that of the control group (P < 0.01). These results suggest that a higher activity of adipogenesis may have been involved in the promoted accumulation of IMF in the L. dorsi muscles of pigs, induced by a dietary LL level.  相似文献   

6.
1. The objective of this study was to estimate the difference between broiler and layer chicks in the activities of calpain and calpastatin (inhibitor of calpain) in breast muscle. Differences between broilers and layers in body weight, daily gain at 3 weeks of age and fractional growth rate (FGR) during 2 and 3 weeks of age were statistically significant (P < 0.01).

2. Calpain and calpastatin activities were measured at three weeks of age with alkali‐denatured casein as a substrate. The m‐calpain (calpain activated by millimolar calcium concentration) activities in units/g muscle and units/ mg extractable muscle protein were 0.779 and 0.353 for broilers, and 1.042 and 0.440 for layers, respectively. The calpastatin activities in units/g muscle and units/mg extractable muscle protein were 0.332 and 0.153 for broilers, and 0.262 and 0.112 for layers, respectively.

3. Broilers with high FGR showed low m‐calpain and high calpastatin activities. In contrast, layers with low FGR showed high m‐calpain and low calpastatin activities.

4. These results suggest that m‐calpain and calpastatin activities in skeletal muscle vary between breeds which have different rates of muscle production.  相似文献   


7.
It has previously been shown that mechanical stretch induces activation of cultured quiescent satellite cells by rapid release of hepatocyte growth factor (HGF) from its extracellular association with satellite cells and its subsequent presentation to the c‐met receptor. The present study provides evidence that the stretch activation activity varies according to the origin of satellite cells from back and leg skeletal muscles in vitro. Satellite cells were isolated from three muscle groups, back (BK), upper hind limb (UL) and lower hind limb (LL) muscles, of adult male rats and stretch activation activities were compared. In response to stretch, lower hind limb satellite cells showed significantly greater response than upper hind limb and back muscles (LL > UL > BK). Immunoblots of stretched culture media revealed a higher HGF‐releasing capacity of lower hind limb satellite cells than back muscle satellite cells. In addition, lower hind limb satellite cells exhibited a greater activation activity in response to exogenous HGF added to culture media than compared to satellite cells from back and upper hind limb (LL > UL > BK). The increased ability to release HGF and the increased cellular responsiveness might account for higher stretch activation activities of lower hind limb satellite cells. Electrophoretic analysis of myosin heavy chain isoforms verified a higher content of slow muscle fibers in lower limb muscles (LL > UL > BK), suggesting a difference in stretch‐induced activation activity between satellite cells associated with fast and slow muscle fibers.  相似文献   

8.
Carcasses from Hanwoo steers (n = 15) and cows (n = 15) were classified into three groups: group 1 (G1), the carcasses had 10% to < 11.5% intramuscular fat (IMF) in loin muscles; group 2 (G2), the carcasses had 13% to < 14.5% IMF in loin muscles; and group 3(G3), the carcasses had 17% to < 18.5% IMF in loin muscles. These were used to evaluate the effects of gender and carcass group on quality traits and Warner–Bratzler shear force (WBSF) of Psoas major (PM), Longissimus thoracis (LT), Longissimus lumborum (LL), Longus colli (LC), Supraspinatus (SS), Latissimus dorsi (LAD), Semimembranosus (SM), Quadriceps femoris (QF), Biceps femoris (BF) and Semitendinosus (ST) muscles. Our results showed that pH values of LT, LL, LC, BF and QF muscles were lower in steers than in cows (P < 0.05). Water holding capacity (WHC) was found higher in LC, SS, LAD and QF muscles of steers (P < 0.05). At day 2 of ageing, gender affected the WBSF values of only PM, LD and QF muscles in G1, and QF muscle in G3; however, with additional ageing, the gender effect was observed for most of the muscles. Most muscles showed ageing responses; however, the rates of ageing response significantly varied depending on gender and carcass groups. The muscles of G1 and G2 had generally higher tenderization potentials than those of G3. Furthermore, most muscles in G3 had generally lower WBSF values than in G1 and G2. These results clearly indicate that ageing has a significant effect on quality and WBSF of beef muscles, and the classification by loin IMF level may be useful for prediction of the tenderness of other muscles.  相似文献   

9.
Calpains are crucial for the degradation of myofibrillar proteins in muscle. Calpastatin is a specific inhibitor of calpains. The objective of this study was to elucidate the effect of nutrient restriction on the activity of calpains and calpastatin in the skeletal muscle of both cows and fetuses. Beginning 30 d after conception, 20 cows were fed either a control diet consisting of native grass hay fortified with vitamins and minerals at recommendations for a mature cow to gain 0.72 kg/d or half the vitamins and minerals and millet straw at 68.1% of NEm requirements. Cows were slaughtered on d 125 of gestation, and the LM was sampled at the 12th rib for calpain and calpastatin measurement. When comparing the muscle samples from nutrient-restricted and control cows, no difference in the activity of calpain I and II was observed; however, there was a significant difference (P < 0.05) in calpastatin activity. Muscle samples from control cows had greater calpastatin content than those of nutrient-restricted cows (P < 0.05); in contrast, the calpastatin content of fetal muscle was greater in fetuses gestated by nutrient-restricted cows than those of control cows (P < 0.05). Further, there were three calpastatin isoforms of 125, 110, and 70 kD detected in fetal muscle, whereas only the110-kD isoform was detected for cow muscle. These results indicate that the activity of the calpain system in skeletal muscle is mainly controlled through the expression of calpastatin. Alternating the calpastatin content in muscle and thereby modulating calpain activity may provide a mechanism for the maintenance of fetal muscle growth during nutrient restriction, whereas skeletal muscle loss in cows is upregulated.  相似文献   

10.
The peroxisome proliferator‐activated receptor‐γ coactivator‐1 α (PGC‐1 α) induces mitochondria biogenesis in skeletal muscles. To determine the relationships between PGC‐1 α and the muscle fiber types, the expression levels of PGC‐1 α were analyzed in porcine and bovine skeletal muscles. As a first step, the nucleotide sequences of the porcine and bovine PGC‐1 α were determined. The porcine and bovine PGC‐1 α cDNA encoded 796 amino acid sequences and showed 95.1% identity between the two species. The expression levels of the PGC‐1 α mRNA were analyzed in the same 10 skeletal muscles from four pigs and three cattle. The contents of porcine and bovine PGC‐1 α were higher in the tongue, masseter and diaphragm, and lower in the Biceps femoris, semimembranosus, Longissimus thoracis and semitendinosus muscles. The contents of myosin heavy chain slow‐type protein (MyHC‐slow) were also determined in the same muscles by ELISA. The analysis of MyHC‐slow showed results similar to those for the PGC‐1 α contents in all of the muscles except for the tongue. The content of MyHC‐slow in the tongue was the lowest among the porcine muscles, and moderate among the bovine muscles. The results suggest that PGC‐1 α relates to the development of oxidative muscle fibers, but is not the principal factor in determining type I fiber content.  相似文献   

11.
This study was conducted to show that dietary supplementation of a fungus, Aspergillus awamori called Koji in Japan, reduces skeletal muscle protein breakdown and stimulates growth in broiler chickens. A total of 30 chicks at 15 days of age was divided into control and two treatment groups (10 birds per treatment). Control group was fed basal diet and treatment groups were fed the basal diets supplemented with A. awamori at levels of 0.05% and 0.2%. The birds were raised for 12 days from 15 to 27 days of age and then the effect on growth, organ weights and plasma 3‐methylhistidine concentration and digestibilities of protein and energy was evaluated. The messenger RNAs (mRNAs) of atrogin‐1, ubiquitin, proteasome, m‐calpain, µ‐calpain, β‐actin, myosin and pax‐7 in the breast muscle were also measured. Body weight gain and breast muscle weight were increased, although feed intake was decreased by the fungus and thus feed efficiency was increased. Protein and energy digestibilities were increased. Furthermore, plasma 3‐methylhistidine concentration was decreased by the fungus. The mRNAs of atrogin‐1, ubiquitin, proteasome, m‐calpain and µ‐calpain were all decreased. The mRNA of β‐actin but not myosin and pax‐7 was slightly increased by the fungus. In conclusion, feeding A. awamori improves growth performance because skeletal muscle proteolytic activity is reduced and digestibilities of energy and protein are increased.  相似文献   

12.
The present study investigated the effects of the nutritional levels of diets on meat quality and related gene expression in Hainan black goat. Twenty‐four goats were divided into six dietary treatments and were fed a concentrate‐based diet with two levels of crude protein (CP) (15% or 17%) and three levels of digestive energy (DE) (11.72, 12.55 or 13.39 MJ/kg DM) for 90 days. Goats fed the concentrate‐based diet with 17% CP had significantly (P < 0.05) higher average daily gains (ADG) and better feed conversion rates (FCR). The pH 24h value tended to decrease (P < 0.05) with increasing DE levels. The tenderness of Longissimus dorsi muscle (LD) and Semimembranosus muscle (SM) reduced with increasing CP levels (P < 0.05). With increasing DE levels, tenderness was increased (P < 0.05). The heart fatty acid‐binding protein (H‐FABP) mRNA expression levels in LD and SM increased with increasing DE levels (P < 0.05), but decreased with increasing CP levels (P < 0.05). The calpastatin (CAST) and μ‐calpain mRNA expressions levels in LD and SM were affected significantly (P < 0.05) by CP and DE levels in the diet. Therefore, the nutritional levels of diets affect meat quality and expression levels of genes associated with meat quality in Hainan black goats.  相似文献   

13.
To assess both quantitative and qualitative differences between the slow‐ and fast‐type muscles, masseter (slow) and semitendinosus (fast) from four Holstein cows were analyzed by two‐dimensional difference gel electrophoresis (2D DIGE) and mass spectrometry. The proteome analysis identified 27 spots as 20 proteins in the whole protein fraction extracted with 8 mol/L urea solution, and 16 spots were identified as 11 proteins in the water‐soluble protein fraction. Two slow‐type myofibrillar proteins (myosin light chain‐1 slow‐b and myosin light chain‐2 slow), and aconitase‐2 mitochondria were present at higher levels in the masseter muscle (P < 0.05). Four fast‐type myofibrillar proteins (myosin light chain‐1 fast, myosin light chain‐2 fast, myosin light chain‐3 fast and tropomyosin‐1), and three enzymes of glycolytic pathway (enolase‐3, aldolase‐A and triosephosphate isomerase), were present at higher levels in the semitendinosus muscle (P < 0.05). Our proteome analysis showed that the composition of sarcoplasmic proteins as well as myofibrillar proteins was clearly different between slow‐ and fast‐type muscles.  相似文献   

14.
Meat tenderness is considered as the most important criterion for meat quality by consumers and can be improved by the actions of endogenous proteases, mainly calpains, during postmortem storage at 0–5°C. The purpose of this study, therefore, was to examine the postmortem calpain activation and proteolysis in breast (BM) and leg and thigh (LM) muscles of White Roman goose. BM and LM were taken from goose carcasses (n = 15) at 0 (10–15 min postmortem), 1, 3, and 7 days of storage at 5°C. The decrease in postmortem pH, calpain‐1 and ‐11 activities, and contents of the calpain‐1 80 kDa subunit and desmin was more rapid (p < .05) in BM than in LM. Our results show that postmortem proteolysis was more extensive in BM than in LM of White Roman goose, not only because the difference in fiber type composition between two muscles, but because the rate and extent of calpain activation were greater in BM as well. These results may provide useful information to optimize meat processing for different muscles in goose industry.  相似文献   

15.
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16.
We developed a ribonuclease protection assay (RPA) to quantify the mRNA mass of calpastatin and the catalytic subunits of calpains I and II in ovine and bovine tissues. The method is based on constructing standard curves using predetermined amounts of in vitro synthesized sense cRNA of the calpain system, hybridized with excess radiolabeled antisense counterprobes. This is possible because the vectors used for riboprobe preparation can be used to transcribe the sense cRNA required to generate the standard curves to quantify absolute calpain I, calpain II, and calpastatin mRNA levels. We used the RPA to study calpain I, calpain II, and calpastatin gene expression in ovine liver, heart, and skeletal muscle. The results revealed that calpain II gene expression was similar in the three tissues. However, the expression of calpain I and calpastatin genes indicates that each tissue has its unique pattern. We also analyzed the activity of calpain I, calpain II, and calpastatin by the conventional DEAE chromatographic method for comparison. The results indicated that the RPA is more repeatable than the DEAE method. Special features of the RPA as compared with DEAE chromatography are as follows; 1) the RPA is a reliable method for quantifying the expression of calpains in all tissues because it is not affected by the presence of inhibitors or activators, 2) the RPA method can be expanded to analyze the expression of the tissue-specific calpains simply by designing specific probes for them, and 3) the RPA requires a small amount of tissue. The described method will facilitate future studies on the gene expression of calpains and will contribute to determining their physiological functions.  相似文献   

17.
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18.
The objectives of this experiment were to assess effects of animal age and castration on activities of calpain I, calpain II, and calpastatin in sheep skeletal muscle. Ten newborn male lambs (2.9 kg), six weaned wethers (23.2 kg), six weaned rams (22.2 kg), six market wethers (55.4 kg), and six market rams (60.2 kg) were slaughtered and samples of biceps femoris were taken for assay of calpain I (micromolar calcium-dependent proteinase), calpain II (millimolar calcium-dependent proteinase), and calpastatin. Preweaning weight gain was similar for rams and wethers; however, postweaning ram growth exceeded (P less than .05) that of wethers. Ram biceps femoris weights at market were greater than those of wethers (P less than .05). Irrespective of age or gender, activity of calpain II was two- to threefold greater than that of calpain I. Muscle calpastatin activity was severa fold higher than calpain I and II activities. Activities of calpains and calpastatin declined (P less than .05) between birth and weaning. A portion of these losses were due to a dilution effect caused by accumulation of muscle proteins. Neonatal attenuation of calpain activities may underlie age-related attenuation of fractional rates of muscle protein degradation. Although ram muscle growth exceeded that of wethers, no differences (P greater than .05) in activities of muscle calpains or calpastatin were detected between these two groups at weaning or at market weight. Hence, castration did not influence lamb muscle growth by altering muscle calpain or calpastatin activities.  相似文献   

19.
This study was conducted to assess the effects of dietary corn oil and vitamin E supplementation on fatty acid (FA) profiles and abundances of acetyl‐CoA carboxylase (ACC) and Δ9 stearoyl‐CoA desaturase (SCD) mRNA of Hu sheep. Animals were allocated to three dietary treatments: basal and supplemented with 3% corn oil (CNO), or CNO plus 500 mg/kg vitamin E (COE). The experiment lasted for 10 weeks. No differences were observed in growth performance and carcass qualities among the three treatments (P > 0.05). Feeding CNO and COE diets increased polyunsaturated FAs including cis 9 trans 11 conjugated linoleic acid, and decreased saturated FA in longissimus muscle (P < 0.05). The mRNA abundances of ACC and SCD as detected by real‐time PCR were reduced (P < 0.05) in liver and subcutaneous fat by supplementary oil, while the SCD mRNA level in longissimus muscle was also reduced (P < 0.05). Inclusion of vitamin E did not have further effects on mRNA abundances of these two enzymes. It is suggested that dietary corn oil supplementation may reduce FA biosynthesis and influence FA profiles in Hu sheep through decreased expression of both ACC and SCD genes.  相似文献   

20.
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.  相似文献   

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