舍饲、放牧对滩羊股二头肌纤维特性及宰后成熟过程中蛋白降解的影响

试验旨在研究舍饲（concentrate feeding,CF）、放牧（pasture feeding,PF）对滩羊股二头肌肉品质、肌纤维特性、宰后成熟过程中超微结构及蛋白降解变化的影响。选取体重接近的4~5月龄滩羊公羔28只,随机分为2组,每组14个重复,每个重复1只羊,分别以舍饲、放牧方式饲养,试验期60 d。试验结束后采集股二头肌作为试验样品,并对其肉质性状、肌纤维特性及宰后成熟过程中超微结构与蛋白降解各项指标进行测定分析。结果显示,放牧组羊肉剪切力、硬度和内聚性均显著高于舍饲组（P<0.05）,弹性则显著低于舍饲组（P<0.05）;放牧组羊肉肌纤维密度和Ⅱ型肌纤维的数量比例均显著低于舍饲组（P<0.05）,Ⅰ型肌纤维的直径、横截面积和数量比例均显著高于舍饲组（P<0.05）。在宰后成熟过程中,舍饲组羊肉肌浆蛋白溶解度显著高于放牧组（P<0.05）;两组肌原纤维蛋白溶解度在宰后24 h至最低后逐渐回升,但各时间点差异均不显著（P>0.05）;肌原纤维小片化指数显著上升后趋于平稳,舍饲组增加率（50.97%）高于放牧组（41.94%）;超微结构显示,宰后肌肉肌原纤维结构松弛、裂解,Z线发生降解,肌原纤维小片化出现,舍饲组肌原纤维结构破坏程度比放牧组更严重。舍饲组肌浆蛋白变性程度小,宰后降解速率较慢,而肌原纤维蛋白降解较明显,降解产生的小分子蛋白量高于放牧组。综上所述,饲养方式改变了滩羊股二头肌纤维特性,并对宰后成熟过程中肌肉超微结构及蛋白降解产生影响,舍饲饲养使得股二头肌纤维密度增加,肌纤维直径与横截面积减小,并降低了肌肉剪切力;此外,由于Ⅱ型肌纤维比例的增加,舍饲饲养提高了肌肉宰后肌原纤维小片化指数增长率、肌浆蛋白溶解度和蛋白降解速率,使宰后成熟过程加快,改善了股二头肌肉嫩度。     

Effect of calcium chloride infusion on the tenderness of lambs fed a beta-adrenergic agonist.

The objective of this study was to examine the effectiveness of CaCl2 infusion in overcoming the toughness of meat associated with dietary administration of a beta- adrenergic agonist (BAA) to lambs. Thirty-two crossbred (1/2 Finnsheep X 1/4 Dorset X 1/4 Rambouillet) wether lambs were randomly assigned to receive 0 or 4 ppm BAA (L644,969; Merck, Sharpe and Dohme Research Laboratories) in a completely mixed, high-concentrate diet for 6 wk. Animals were slaughtered in two groups of 16. At each slaughter time half of each group (0 or 4 ppm BAA) was randomly assigned to CaCl2 infusion. Feeding the BAA decreased (P less than .05) fat thickness, kidney-pelvic fat, yield grade, and marbling and increased (P less than .05) dressing percentage, lean firmness, leg score, and biceps femoris weight. Weight of biceps femoris was 32.8% greater in BAA-fed lambs. Treated, but not infused, lambs were significantly less tender than control lambs after 1, 7, and 14 d of postmortem storage. At 24 h postmortem, BAA-fed lambs had higher (P less than .05) cathepsin B, calcium-dependent protease-II (CDP-II), and CDP inhibitor activities. Calcium chloride infusion increased marbling, decreased lean firmness, increased lean color score, and increased dressing percentage (P less than .05). Infusion of carcasses with CaCl2 decreased (P less than .05) shear force at all postmortem times. Infusion of carcasses with CaCl2 had no effect on cathepsins B and B + L activities, but it had a significant effect on CDP-I, CDP-II, and CDP inhibitor activities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Postmortem proteolysis and calpain/calpastatin activity in callipyge and normal lamb biceps femoris during extended postmortem storage.

The present experiment was conducted to determine whether calpastatin inhibits only the rate, or both the rate and extent, of calpain-induced postmortem proteolysis. Biceps femoris from normal (n = 6) and callipyge (n = 6) lamb was stored for 56 d at 4 degrees C. Calpastatin activity was higher (P < .05) in the callipyge muscle at 0 and 14 d postmortem, but not at 56 d postmortem. The activity of mu-calpain did not differ between normal and callipyge biceps femoris at 0 and 56 d postmortem (P > .05), but was higher at 14 d postmortem in the callipyge muscle (P < 0.05). The activity of m-calpain was higher in the callipyge muscle (P < 0.05). Western blot analyses of titin, nebulin, dystrophin, myosin heavy chain, vinculin, alpha-actinin, desmin, and troponin-T indicated that postmortem proteolysis was less extensive in callipyge than in normal biceps femoris at all postmortem times. The results of this experiment indicate that calpastatin inhibits both the rate and extent of postmortem proteolysis.     

Reduced calcium-dependent proteinase activity in cimaterol-induced muscle hypertrophy in lambs

The purpose of this study was to reveal the possible relationships between total Ca2+-dependent proteinase (CDP) and micromolar-Ca2+-dependent proteinase (microM CDP) activity and cimaterol-induced hypertrophy of skeletal muscle. Dietary administration of 10 ppm cimaterol to finishing lambs reduced microM CDP activity in longissimus muscle (LD) by 55% (P less than .01) and 70% (P less than .02) after 3 and 6 wk of treatment, respectively. Total CDP activity was unaffected by cimaterol at both treatment intervals. The reduced microM CDP activity was not associated with a reduced yield of enzyme extract from the muscle. Cimaterol treatment increased the cross-sectional area of the LD by 23.5% at 3 wk and by 35.6% at 6 wk (P less than .001). Cimaterol also increased (P less than .001) the masses of semitendinosus, semimembranosus and biceps femoris by 26%, 32.4% and 24.5%, respectively. These results suggest that cimaterol-induced muscle hypertrophy may be attained in part by reduction of myofibrillar protein degradation.     

The calpain system in three muscles of normal and callipyge sheep

Activities of mu- and m-calpain and of calpastatin were measured at four different times during postmortem storage (0, 1, 3, and 10 d) in three muscles from either callipyge or noncallipyge (normal) sheep. The weights of two muscles, the biceps femoris and the longissimus, are greater in the callipyge phenotype, whereas the weight of the infraspinatus is not affected. The activity of m-calpain was greater (P < 0.05) in the biceps femoris and longissimus from callipyge than in those from normal sheep, but it was the same in the infraspinatus in the two phenotypes. The extractable activity of m-calpain did not change (biceps femoris and infraspinatus) or decreased slightly (longissimus) during postmortem storage. Extractable activity of mu-calpain decreased to zero or nearly zero after 10 d postmortem in all muscles from both groups of sheep. The rate of decrease in mu-calpain activity was the same in muscles from the callipyge and normal sheep. At all time points during postmortem storage, calpastatin activity was greater (P < 0.05) in the biceps femoris and longissimus from the callipyge than from the normal sheep, but it was the same in the infraspinatus from callipyge and normal sheep. Calpastatin activity decreased (P < 0.05) in all three muscles from both phenotypes during postmortem storage; the rate of this decrease in the callipyge biceps femoris and longissimus and in the infraspinatus from both the callipyge and normal sheep was slow, especially after the first 24 h postmortem, whereas calpastatin activity in the biceps femoris and longissimus from the normal sheep decreased rapidly. During postmortem storage, the 125-kDa calpastatin polypeptide was degraded, but the 80-kDa subunit of mu-calpain was cleaved only to 76- and 78-kDa polypeptides even though extractable mu-calpain activity declined nearly to zero. Approximately 50 to 60% of total mu-calpain became associated with the nonextractable pellet after 1 d postmortem. The myofibril fragmentation index for the biceps femoris and longissimus from normal sheep increased significantly during postmortem storage. The fragmentation index for the infraspinatus from the callipyge and normal sheep increased to an intermediate extent, whereas the index for the biceps femoris and longissimus from the callipyge did not change during 10-d postmortem storage. The results suggest that postmortem tenderization is related to the rate of calpastatin degradation in postmortem muscle and that calpastatin inhibition of the calpains in postmortem muscle is modulated in some as yet unknown manner.     

Cimaterol-induced muscle hypertrophy and altered endocrine status in lambs

The objectives of this study were 1) to determine how cellular growth of skeletal muscle is altered by the repartitioning agent cimaterol and 2) to determine if cimaterol alters endocrine status in association with its repartitioning effects. Thirty Dorset wether lambs were randomly assigned to a pre-treatment baseline group or received 0 or 10 ppm cimaterol in a complete, mixed, high-concentrate diet for 7 or 12 wk. Weights of biceps femoris (BF), semimembranosus (SM) and semitendinosus (ST) muscles were 32.8, 27.1 and 31.5% greater, respectively, in treated lambs at 7 wk, and were 22 to 24% greater at 12 wk. Longissimus (LD) cross-sectional area was 26 and 32% greater at these treatment intervals. Percent type I fibers declined significantly over the course of the experiment in ST, SM and LD, and cimaterol caused a small but significant reduction in percent type I fibers in the ST at 7 and 12 wk. Muscles from lambs fed cimaterol contained 50 and 75% more fibers that exhibited negative staining for phosphorylase activity. Mean cross-sectional area of type I and type II fibers in the combined portions of the ST were 30.4 and 29.3% greater, respectively, in cimaterol-fed lambs after 12 wk, while type I and type II fiber areas in the longissimus were only 13 and 15% greater, respectively. Cimaterol-induced hypertrophy of the ST resulted in both protein and RNA content being 30 to 35% greater (P less than .01) at 7 and 12 wk, while DNA concentration was 22% less (P less than .01) at 7 wk. DNA concentration returned to normal by 12 wk. These results indicate that cimaterol elicits a rapid increase in muscle RNA and protein accretion without concurrent incorporation of satellite cell nuclei. Plasma insulin and insulin-like growth factor-1 (IGF-1) concentrations were 55 and 34% lower, respectively, in cimaterol-fed lambs. Plasma somatotropin concentration and area under the curve were 2.3 times greater (P less than .01) in lambs fed cimaterol for 6 wk, while plasma cortisol, prolactin and glucose concentration were unaffected at 6 or 12 wk. The significant changes in endocrine status may be important in the mechanism(s) of cimaterol in altering muscle accretion.     

饲养方式对滩羊羔羊肌肉及脂肪组织中脂肪酸组成的影响

为探究饲养方式对滩羊羔羊肌肉和脂肪组织中脂肪酸组成的影响,试验选择24只健康且体重相近（14.16 kg±0.58 kg）的2月龄滩羊羔羊随机分成舍饲组和放牧组,每组12只,于4月龄屠宰取其背最长肌、股二头肌、膈肌、皮下脂肪、肾周脂肪、尾部脂肪,用气相色谱仪测定脂肪酸含量。结果表明：①在肌肉组织饱和脂肪酸中,与放牧组相比,舍饲组滩羊羔羊股二头肌丁酸、棕榈酸含量显著提高（P<0.05）,膈肌辛酸、月桂酸、肉豆蔻酸含量显著降低（P<0.05）。在肌肉组织不饱和脂肪酸中,与放牧组比较,舍饲组滩羊羔羊背最长肌棕榈油酸、花生四烯酸含量显著降低（P<0.05）,γ-亚麻酸、α-亚麻酸含量显著提高（P<0.05）,二十碳五烯酸（EPA）、n-3多不饱和脂肪酸（n-3 PUFA）极显著提高（P<0.01）;股二头肌花生四烯酸含量显著降低（P<0.05）,十七碳一烯酸、α-亚麻酸、EPA、n-3 PUFA含量极显著提高（P<0.01）;膈肌棕榈油酸含量显著降低（P<0.05）。②在脂肪组织饱和脂肪酸中,与放牧组比较,舍饲滩羊羔羊皮下脂肪十一碳酸、十五碳酸含量显著降低（P<0.05）,十三碳酸含量极显著降低（P<0.01）;肾周脂肪葵酸、肉豆蔻酸含量显著降低（P<0.05）,丁酸、硬脂酸含量极显著提高（P<0.01）;尾脂十一碳酸含量显著降低（P<0.05）。在脂肪组织不饱和脂肪酸中,与放牧组比较,舍饲滩羊羔羊皮下脂肪十八碳二烯酸含量极显著提高（P<0.01）,肉豆蔻油酸、棕榈油酸、γ-亚麻酸含量极显著降低（P<0.01）;肾周脂肪十八碳一烯酸、α-亚麻酸、n-3PUFA含量显著提高（P<0.05）,肉豆蔻油酸含量显著降低（P<0.05）,十七碳一烯酸、二十碳二烯酸、十八碳二烯酸含量极显著提高（P<0.01）;尾脂肉豆蔻油酸、棕榈油酸含量显著降低（P<0.05）,十八碳二烯酸含量极显著提高（P<0.01）。综上,不同饲养方式对滩羊羔羊肌肉和脂肪组织脂肪酸含量存在一定影响,舍饲滩羊羔羊股二头肌中饱和脂肪酸含量及肾周脂肪中不饱和脂肪酸含量显著高于放牧滩羊羔羊。     

Alterations in postmortem degradation of myofibrillar proteins in muscle of lambs fed a beta-adrenergic agonist

Dietary administration of 4 ppm of the beta-agonist L-644,969 (Merck Sharpe and Dohme Research Laboratories) to finishing lambs induced a decrease (10 to 14%, P less than .05) in extractable calpain I activity in the longissimus muscle (LD) at death (d 0). At 4 d postmortem (d 4), extractable calpain I levels in the LD of both control and treated lambs were reduced (P less than .001) from those present at d 0, but the extractable activity in the LD was reduced to a greater extent in control than in treated lambs. Calpain II activity was increased 42% (P less than .005) in LD of treated lambs; however, no significant differences were observed between d 0 and d 4 calpain II activity within treated or control LD samples (P greater than .1). Calpastatin activity was higher in the LD of treated lambs (74% on d 0, P less than .001 and 430% on d 4, P less than .001) than in the LD of control lambs. Measurable cathepsin B activity was decreased (29% on d 0, P less than .05) and measurable cathepsin H activity was increased (10% on d 0, P less than .05 and 10% on d 4, P less than .05) in the LD of treated lambs compared with controls. On d 2, 4 and 6 postmortem, degradation in myofibrils isolated from the LD was lower for treated than for control lambs. Warner-Bratzler shear values for loin chops from treated lambs were higher on both d 3 (111%) and 6 (108%) postmortem than for chops from control lambs (P less than .001). L-644,969-induced decreases in muscle proteolytic capacity may limit postmortem myofibril degradation and contribute to the reduced tenderness observed. This decreased proteolytic capacity may contribute to increased muscularity of L-644,969-treated lambs.     

Variation in proteolysis, sarcomere length, collagen content, and tenderness among major pork muscles

The objectives of this experiment were to determine the extent of variation in proteolysis, sarcomere length, and collagen content among pork muscles and the association of those factors with tenderness variation among muscles at 1 d postmortem. Twenty-three white composite barrows were slaughtered and carcasses (66 kg) were chilled at 0 degrees C for 24 h. At 1 d postmortem, the longissimus lumborum, biceps femoris, semimembranosus, semitendinosus, and triceps brachii, long head were dissected from one side of each carcass and frozen. Trained sensory panelists evaluated tenderness, amount of connective tissue, juiciness, and pork flavor intensity of grilled (70 degrees C) chops on 8-point scales. Raw chops were used for total collagen content, sarcomere length, and the extent of desmin proteolysis. Tenderness ratings were highest (P < .05) for semitendinosus (7.2) and triceps brachii (7.1), followed by longissimus lumborum (6.4) and semimembranosus (5.7) and were lowest (P < .05) for biceps femorus (4.0). The simple correlations between longissimus lumborum tenderness and the tenderness of other muscles were .54 (semimembranosus), .34 (semitendinosus), .36 (triceps branchii), and .17 (biceps femorus). Total collagen was highest (P < .05) for biceps femorus (7.1 mg/g muscle), followed by triceps branchii (6.0 mg/g) and semitendinosus (5.3 mg/g), and lowest for semimembranosus (4.5 mg/g) and longissimus lumborum (4.1 mg/g). Sarcomere length was longest (P < .05) for semitendinosus (2.5 microm) and triceps branchii (2.4 microm), followed by semimembranosus (1.8 microm), longissimus lumborum (1.8 microm), and biceps femorus (1.7 microm). Proteolysis of desmin was greatest (P < .05) in longissimus lumborum (39.3%), followed by semimembranosus (21.0%) and biceps femoris (18.5%), then semitendinosus (.2%) and triceps brachii (.2%). Multiple linear regression using total collagen, sarcomere length, and proteolysis accounted for 57% of the variation in tenderness rating among all samples. Piecewise linear regression was used to account for the interaction of sarcomere length with proteolysis and collagen. This analysis accounted for 72% of the variation in tenderness rating. Variation in collagen, proteolysis, and sarcomere length and the degree of their interaction with one another determine the tenderness of individual muscles.     

Isoelectric focusing under dissociating conditions for analysis of muscle protein from clinically normal dogs and Labrador retrievers with hereditary myopathy

Protein profiles of whole homogenates of anconeus (slow twitch) and biceps femoris (fast twitch) muscles of clinically normal dogs and of Labrador Retrievers with hereditary myopathy (HM) were resolved on flat bed polyacrylamide isoelectric-focusing gels. Three methods of sample solubilization were performed. The solubilization buffer, with high concentrations of urea, precipitated the zwitterionic detergent, but use of the buffer containing 3% NP-40, 9.2M urea, and 0.1M arginine resulted in better resolution and stability of pH gradient. Gels of anconeus muscle from clinically normal dogs contained 2 protein bands specific to anconeus muscle, whereas gels of biceps femoris muscle from clinically normal dogs contained 3 protein bands amplified in biceps femoris muscle that were barely detectable in anconeus muscle. The staining intensity of protein bands in biceps femoris muscles from Labrador Retrievers with HM was decreased, relative to controls. The quantitative analysis of peak height ratios of biceps femoris muscle revealed significant (P less than 0.05) differences between profiles of clinically normal dogs and Labrador Retrievers with HM.     

Effects of age and castration on activities of calpains and calpastatin in sheep skeletal muscle.

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.     

湖羊肌肉营养特点及肌纤维组织学特性

为了研究湖羊肌肉营养特点及肌纤维组织学特性,分别测定了不同年龄(初生、断奶、成年)湖羊肌肉中常规营养成分(水分、粗蛋白质、粗脂肪、粗灰分、钙、磷)、18种氨基酸的含量以及不同年龄湖羊不同部位肌肉(臂三头肌、股二头肌、背最长肌)的肌纤维组织学特性(肌纤维直径、肌纤维密度)。结果表明:随着年龄的增长,肌肉中水分含量显著降低(P0.05),粗脂肪、粗蛋白质、粗灰分、钙和磷的含量逐渐增加,且成年羊与初生羔羊和断奶羔羊之间差异显著(P0.05)。湖羊肌肉中苏氨酸、缬氨酸、蛋氨酸、异亮氨酸、亮氨酸、苯丙氨酸、赖氨酸、组氨酸和精氨酸的含量随着年龄的增长而显著增加(P0.05),非必需氨基酸含量随着年龄的增长大致呈增加趋势。在湖羊肌肉的所有氨基酸中含量最高的是谷氨酸,其次是天冬氨酸。不同年龄湖羊的肌纤维直径均表现为背最长肌股二头肌臂三头肌,肌纤维密度均表现为背最长肌股二头肌臂三头肌,且同龄湖羊3个部位之间的肌纤维密度均差异显著(P0.05)。相同部位的肌肉,其肌纤维直径随年龄的增长而增大,肌纤维密度随年龄的增长而减小,且不同年龄间差异显著(初生羔羊与断奶羔羊的臂三头肌除外)(P0.05)。综上可知,湖羊肌肉营养丰富,氨基酸种类齐全,肌纤维组织学特性在不同年龄和不同部位间存在差异,断奶羔羊肉营养更丰富,更具香、嫩等特点。     

Effects of cimaterol on rabbit growth and myofibrillar protein degradation and on calcium-dependent proteinase and calpastatin activities in skeletal muscle

The objectives of this study were to examine effects of a beta-adrenergic agonist (cimaterol) on growth and muscle development in rabbits and to examine cimaterol's effects on myofibrillar protein degradation (MPD) and on activities of several proteolytic enzymes including the calcium-dependent proteinases (CDP). Twelve New Zealand White rabbits were assigned to either control diets or to diets containing cimaterol for 35 d, after which they were killed and effects on performance and tissue weight gains were determined. Urine was collected from d 21 through 28 from each rabbit for assessment of N tau-methylhistidine (NMH) excretion. Cimaterol increased rates of gain, efficiency of gain and skeletal muscle weights. Enhancement in muscle weight was associated with an increase in total DNA and with a reduction in NMH. Cimaterol did not affect activities of cathepsin B, cathepsin D or neutral serine proteinase, but it reduced activities of the millimolar and micromolar forms of the CDP by 58 and 57%, respectively, and it reduced activity of the inhibitor of the CDP (calpastatin) by 52%. Cimaterol-dependent myofibrillar protein accretion was likely mediated, at least in part, by a reduction in MPD. The change in MPD was associated with a reduction in muscle CDP activities. Cimaterol-dependent muscle hypertrophy therefore may involve changes in calcium-dependent proteolysis of myofibrillar proteins. The significance of the effects of cimaterol on calpastatin activity is not known.     

Effects of manipulation of the caspase system on myofibrillar protein degradation in vitro

Apoptosis via the intrinsic caspase 9 pathway can be induced by oxidative stressors hydrogen peroxide (H?O?) and N-(4 hydroxyphenol) rentinamide (fenretinide), a synthetic retinoid. Accelerated muscle atrophy and proteolysis in muscle-wasting conditions have been linked to oxidative stress and activated protease systems. Therefore, the hypothesis of this study was that proteolysis of myofibrillar proteins could be manipulated through the induction or inhibition of the caspase system. After slaughter, LM and supraspinatus muscles from callipyge (n = 5) and normal (n = 3) lambs were excised, finely diced, and incubated with treatment buffers containing oxidative stressors fenretinide or H?O?, recombinant caspase 3, caspase-specific inhibitor N-acetyl-Asp-Glu-Val-Asp-CHO (DEVD), or control solution. Muscle samples were incubated for 1, 2, 7, and 21 d at 4°C. Activation of the initiator caspase, caspase 9, and myofibrillar protein degradation was determined by SDS-PAGE and Western blotting. Results showed that fenretinide, H?O?, and recombinant caspase 3 increased (P < 0.05) proteolysis of myofibril proteins, whereas DEVD inhibited degradation (P < 0.05). Proteolysis of myofibrillar proteins increased with incubation time (P < 0.0001), and incubation time × treatment interactions (P < 0.05) indicated that the treatment effects did not all occur at the same rate. This study has shown that manipulation of the caspase system through induction or inhibition of activity can affect degradation of myofibrillar proteins, providing further evidence that the caspase system could be involved in postmortem proteolysis and tenderization. However, these stimulated changes were not sufficient to overcome the lack of proteolysis that is characteristic of muscle from callipyge lambs.     

Protein kinetics in callipyge lambs

The objectives for this experiment were to determine the effect of the callipyge phenotype on protein kinetics. We studied callipyge and normal lambs (n = 37) at 5, 8, and 11 wk of age (n = 4 to 7/ group) to determine how protein kinetics are altered by this trait. Total protein, DNA, and RNA and calpastatin activity were measured in five skeletal muscles and in the heart, kidneys, and liver, and protein accretion rates were calculated. At 8 wk, the fractional synthesis rates of proteins in these tissues were measured in vivo using a primed, continuous 8-h infusion of [2H5]phenylalanine. Fractional rates of protein degradation were estimated by differences. At 5 wk of age, muscle weights, protein mass, protein:DNA, RNA:DNA, and calpastatin activity were higher (P < .05) for callipyge, and protein mass differences continued to increase through 11 wk. At 8 wk, fractional rates of protein synthesis and degradation were lower (P < .05) in callipyge than in normal lambs. The organs of callipyge lambs exhibited reduced growth at 11 wk. Thus, enhanced muscle growth seems to be maintained in callipyge lambs by reduced protein degradation rather than increased protein synthesis. However, we cannot exclude the possibility that the initial onset of the callipyge condition may be caused by an increase in the fractional rate of protein synthesis.     

Effects of birth weight and postnatal nutrition on neonatal sheep: II. Skeletal muscle growth and development

This study investigated effects of birth weight and postnatal nutrition on growth and development of skeletal muscles in neonatal lambs. Low (L; mean +/- SD 2.289 +/- .341 kg, n = 28) and high (H; 4.840 +/- .446 kg, n = 20) birth weight male Suffolk x (Finnsheep x Dorset) lambs were individually reared on a liquid diet to grow rapidly (ad libitum fed, ADG 337 g, n = 20) or slowly (ADG 150 g, n = 20) from birth to live weights (LW) up to approximately 20 kg. At birth, weight of semitendinosus (ST) muscle in L lambs was 43% that in H lambs; aggregate weights of ST and seven other dissected muscles were similarly reduced. In ST muscle of L lambs, mass of DNA, RNA, and protein were also significantly reduced to levels 67, 60, and 34%, respectively, of those in H lambs. However, myofiber numbers of ST, tibialis caudalis, or soleus muscles did not differ between the L and H birth weight lambs and did not change during postnatal growth. During postnatal rearing, daily accretion rate of dissected muscle was lower in L than in H lambs. Accretion of muscle per kilogram of gain in empty body weight (EBW) was reduced in the slowly grown L lambs compared with their H counterparts, although the difference was less pronounced between the rapidly grown L and H lambs. Throughout the postnatal growth period, ST muscle of L lambs contained less DNA with a higher protein:DNA ratio at any given muscle weight than that of H lambs. Slowly grown lambs had heavier muscles at any given EBW than rapidly grown lambs. Content of DNA and protein:DNA ratio in ST muscle were unaffected by postnatal nutrition, but RNA content and RNA:DNA were greater and protein:RNA was lower at any given muscle weight in rapidly grown lambs. Results suggest that myofiber number in fetal sheep muscles is established before the presumed, negative effects of inadequate fetal nutrient supply on skeletal muscle growth and development become apparent. However, proliferation of myonuclei may be influenced by fetal nutrition in late pregnancy. Reduced myonuclei number in severely growth-retarded newborn lambs may limit the capacity for postnatal growth of skeletal muscles.     

Characteristics of Lacha and Rasa Aragonesa lambs slaughtered at three live weights

A study was made of differences in the quality of meat from Lacha (L) and Rasa Aragonesa (RA) lambs slaughtered at 12, 24, or 36 kg live weight. Lambs from both breeds were weaned at 25 to 57 d, approximately 11.5 to 18.5 kg live weight, and fed concentrate and barley straw until slaughter at 24 and 36 kg live weight. Hot carcass weight, cold carcass weight, conformation, color, firmness, and thickness of backfat and color of rectus abdominis muscle were recorded on the carcass. Final pH (pHu), instrumental color (L*, a*, b*), myoglobin concentration, chemical composition, and water-holding capacity (WHC) of the longissimus muscle, shear force of the biceps femoris muscle, and iodine values and fatty acid composition of the i.m. and s.c. fat depots were determined. The percentage of fat in the longissimus muscle increased with live weight, and values for RA lambs were higher than those for L lambs. The WHC of meat from RA lambs was lower at 24 kg than at 12 or 36 kg slaughter weight. Live weight and breed had no effect on the shear force of the biceps femoris muscle. There was an increase in myoglobin concentration in the longissimus muscle with increased live weight in both breeds. The fatty acid content of s.c. and i.m. fat, which was not affected by breed, declined with the increase in slaughter weight. The polyunsaturated fatty acid content of the s.c. fat depot increased, whereas that of the i.m. fat depot decreased, with the increase in slaughter weight in both breeds. Subcutaneous fat had a higher content of heptadecanoic acid (17:0) than i.m. fat, and this increased with the increase in slaughter weight. In both depots, there was an increase in oleic acid (18:1) at 12 kg in RA lambs and at 24 kg in L lambs. In the s.c. fat depot, there was a progressive increase in linoleic acid (18:2) content with the increase in live weight in both breeds. There was a higher degree of unsaturation in the s.c. fat of RA lambs than in that of L lambs, which was reflected in the iodine value.     

Elevated rate of collagen solubilization and postmortem degradation in muscles of lambs with high growth rates: possible relationship with activity of matrix metalloproteinases

The extracellular matrix, composed mainly of collagen, is considered responsible for the residual toughness of meat. Matrix metalloproteinases (MMP) responsible for the degradation of connective tissue are found in most tissues, but their participation in meat aging has not been tested. We recently showed that skeletal muscle has multiple MMP activities, as well as regulators and tissue inhibitors of metalloproteinases. Here we present the first observations of physiologic and postmortem variation of MMP activities in muscle. Growing lambs were offered two levels of intake: hay + concentrate for lambs with high growth rate (average daily gain > 250 g) and hay only for those with low growth rate (average daily gain < 25 g). At slaughter and at 21 d of postmortem aging of longissimus and semimembranosus muscles, we studied collagen content, collagen solubility, free hydroxyproline (OH-pro), and levels of latent and active forms of a matrix metalloproteinase (MMP-2) by gelatin zymography. Our results demonstrate the presence of an active isoform of MMP-2 in lamb muscle. Its level was higher (+90%, P < 0.01) in lambs that expressed a high growth rate. Activity of MMP-2 was also present at 21 d postmortem, at levels similar to those detected at slaughter. At slaughter and at 21 d, all muscles contained latent MMP-2 and the quantity of proenzyme was greater than that present in the activated form. The levels of free OH-pro in muscles of lambs with high growth rate increased significantly (P < 0.001) over 21 d from 3.75 to 5.08% of total collagen, and this was significantly related to the level of active MMP-2 at slaughter. By contrast, the amount of free OH-pro in muscles of lambs with low growth rate was not different at 21 d (1.63% of total OH-pro) than it had been at slaughter (1.84% of total OH-pro). These results suggest that collagen degradation all the way to free amino acids occurs postmortem in muscle and that there are active MMP simultaneously present that may account for this catabolism. The growth rate of animals at slaughter influences collagen turnover in vivo, as well as postmortem collagen degradation.     

Postmortem proteolysis is reduced in transgenic mice overexpressing calpastatin

Using both in vitro and in vivo approaches, numerous studies have provided evidence that mu-calpain is responsible for postmortem proteolysis. This paper reports the effect of overexpression of calpastatin on postmortem proteolysis in transgenic mice. Transgenic mice (n = 8) with a human calpastatin gene, whose expression was driven by the human skeletal muscle actin promoter, were killed along with control nontransgenic littermates (n = 5). Hind limbs were removed and stored at 4 degrees C, and muscle samples were dissected at 0, 1, 3, and 7 d postmortem and analyzed individually. At time 0, active human calpastatin was expressed in transgenic murine skeletal muscle at a level 370-fold greater (P < 0.001) than calpastatin in control mice. Although the native isoform of this protein was degraded with storage, at 7 d postmortem, approximately 78% of at-death activity remained, indicating that degraded calpastatin retains activity. Calpain (mu- and m-) expression was unaffected (P > 0.05) by the transgene as assessed by immunoreactivity at d 0. Over 7 d, 33% of at-death 80-kDa isoform immunoreactivity of mu-calpain was lost in transgenics compared to an 87% loss in controls, indicating that autolysis of mu-calpain was slowed in transgenic mice. Desmin degradation was also inhibited (P < 0.05) in transgenics when compared to controls. Control mice lost 6, 78, and 91% of at-death native desmin at 1, 3, and 7 d postmortem, respectively; conversely, transgenic mice lost only 1, 3, and 17% at the same times. A similar trend was observed when examining the degradation of troponin-T. Interestingly, m-calpain seemed to undergo autolysis in control mice, which in postmortem tissue is indicative of proteolysis. Further investigation revealed that both mu- and m-calpain are active postmortem in normal murine skeletal muscle. In conclusion, a high level of expression of active calpastatin was achieved, which, by virtue of its inhibitory specificity, was determined to be directly responsible for a decrease in postmortem proteolysis.     

Feeding high levels of vitamin D3 does not improve tenderness of callipyge lamb loin chops

The objective of this study was to determine whether feeding high doses of vitamin D3 7 d before slaughter would increase muscle Ca++ levels and result in more tender loin chops. Market lambs (n = 4 callipyge and 4 normal in Exp. 1, and n = 16 calipyge and 16 normal in Exp. 2) were randomly and equally assigned to feeding groups based on callipyge genotype and experimental diet, (vitamin D3 or control). Serum Ca++, muscle Ca++, Warner-Bratzler shear force, and troponin-T degradation data were analyzed. In Exp. 1, vitamin D3 was supplemented at 1 or 2 x 10(6) IU/d. The 2 x 10(6) IU dose resulted in the greatest serum Ca++ reponse and was chosen for Exp. 2. In Exp. 2, serum Ca++ concentration was higher (P < 0.05) for normal and callipyge lambs fed the vitamin D3 diet than for the control diet fed lambs. Muscle Ca++ concentrations, however, were not higher (P = 0.28) for the vitamin D3-fed lambs. Warner-Bratzler shear values were higher (P < 0.05) for callipyge than for normal lambs, but no differences were observed with vitamin D3 supplementation. These data were supported by results from Western blot analysis of troponin-T degradation, in which no differences were observed for vitamin D3 vs control diet lambs at 14 d postmortem. This experiment showed that feeding 2 x 10(6) IU/d of vitamin D3 to market lambs, callipyge or normal, raised serum Ca++ concentration, but did not increase muscle Ca++ concentration. This lack of response in muscle Ca++ was likely the reason that no differences were observed for Warner-Bratzler shear force values or troponin-T degradation data between the vitamin D3 and control loin chops. A higher dose of vitamin D3 may be required to improve tenderness.