Relationships between tropomyosin and myosin heavy chain isoforms in bovine skeletal muscle

The composition of tropomyosin (TPM) and myosin heavy chain (MyHC) isoforms was analyzed in 10 physiologically different bovine muscles ( masseter , diaphragm, tongue, semispinalis, pectoralis profundus , biceps femoris, psoas major , semimembranosus, longissimus thoracis and semitendinosus ) to clarify the relationships between TPM and MyHC isoforms in different muscle fiber types. The content of TPM1 and TPM3 was different in muscles according to their function in muscle contraction, although the content of TPM2 was constantly about 50% of the total TPM in all muscles. The content of TPM1 was higher in semimembranosus , longissimus thoracis and semitendinosus, while that of TPM3 was higher in masseter and diaphragm. The high positive correlation between MyHC-slow content and TPM3 content ( r  = 0.92) suggested a coexpression of TPM3 and MyHC-slow isoforms in a muscle fiber. MyHC-slow and TPM3 were expressed at the same level in masseter and diaphragm, whereas there was more TPM3 than MyHC-slow in tongue and semispinalis , so it appears that the excess TPM3 in tongue and semispinalis is expressed with other MyHC isoforms. MyHC-2a was the only fast type isoform expressed in tongue and semispinalis . Therefore, the excess TPM3 was composed of myofibrils with MyHC-2a. The results suggested that a fiber expressing MyHC-2a would be regulated delicately by changing the TPM isoform types.     

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.     

Peroxisome proliferator-activated receptor-γ coactivator 1 α (PGC-1 α) expression and the formation of slow-twitch muscle fibers in porcine and bovine skeletal muscles

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.     

Amino acid sequences of multiple fast and slow troponin T isoforms expressed in adult bovine skeletal muscles

Multiple nucleotide sequences of complementary DNA (cDNA) of bovine troponin T (TnT) isoforms expressed in the adult skeletal muscles were determined to facilitate the elucidation of the TnT degradation progress during postmortem aging of muscles. Fresh muscle samples were excised from the lingual, masseter, pectoralis, diaphragm, psoas major, longissimus thoracis, spinnalis, semitendinosus, semimembranosus, and biceps femoris muscles of three Holstein cows within 1 h of slaughter. Complementary DNA fragments of fast and slow TnT isoforms expressed in each muscle were amplified by reverse-transcribed PCR. Consequently, four major fragments of fast TnT and two fragments of slow TnT, all of which contained the complete coding region, were obtained. The sequence determination of these fragments revealed that at least eight and two isoforms were generated by the alternative splicing from bovine fast and slow TnT messenger RNA, respectively. In the fast TnT isoforms, five small variable exons were observed; three of these five exons were in the amino (N)-terminal region. The calculated molecular weight of fast and slow TnT isoforms ranged from 29,816 to 32,125 and from 30,166 to 31,284, respectively. The deduced amino acid sequences revealed that the N-terminal region of all the TnT isoforms was extremely glutamic acid-rich. Reverse-transcribed PCR analysis revealed that expression of each of these isoforms was distributed in a fast or slow muscle-specific manner. Given that TnT degradation has been reported to accompany a decrease in glutamic acid content in the conventional 30-kDa degradation product, the sequence data suggested that the 30-kDa fragment seem to be generated by the proteolytic removal of the glutamic acid-rich N-terminal ends. The multiplicity of TnT isoforms may result in a complicated pattern of TnT degradation on SDS-PAGE gel during beef aging.     

Enhanced myotoxicity and involvement of both type I and II fibers in monensin-tiamulin toxicosis in pigs

Simultaneous administration of monensin and tiamulin to pigs resulted in enhanced myotoxicity. Skeletal muscles of tongue, diaphragm and legs were preferentially affected, whereas the masseter, longissimus thoracis and cardiac muscles, including the left auricle, were spared. Histochemical examination revealed an involvement of both type I and II fibers of skeletal muscles.     

中国西门塔尔牛肌肉肌纤维类型和肉品质特性研究

为研究中国西门塔尔牛的肌纤维和肉品质特性,本试验以10头中国西门塔尔公牛为研究对象,测定了背最长肌、腰大肌和半腱肌的组织学特性、pH、系水力、剪切力和质构剖面分析。结果显示,肌纤维直径Ⅰ型 < ⅡA型 < ⅡB型,腰大肌中Ⅰ型具有最高的肌纤维数比例和肌纤维面积比,背最长肌和半腱肌中ⅡB型具有最高的肌纤维比例和肌纤维面积比。与腰大肌相比,背最长肌和半腱肌具有较低pH、弹性和压力失水率,较高的剪切力、硬度和回复性。综上所述,肌纤维特性影响牛肉品质,特别是牛肉的质构特性,提高Ⅰ型纤维比例和降低ⅡB型纤维比例能够改善牛肉嫩度。     

Study on Muscle Fiber Types and Meat Quality Traits of Chinese Simmental Cattle

The aim of this test was to study muscle fiber characteristics and meat quality traits of Chinese Simmental cattle.Ten Chinese Simmental bulls were selected and longissimus dorsi,psoas major and semitendinosus were collected.Histological characteristics,pH,water holding capacity,shear force and texture profile analysis (TPA) were measured.The results showed that the diameter of these muscle fibers increased in the rank order type Ⅰ < ⅡA < ⅡB.The number and area percentage of type Ⅰ muscle fiber were highest in longissimus dorsi and that of ⅡB muscle fiber was highest in psoas major and semitendinosus.Longissimus dorsi and semitendinosus had lower pH,springiness,pressing lossing rate and higher shear force,hardness and resilience comparing with psoas major.In conclusion,muscle fiber characteristics could affect meat quality,especially textural properties of Chinese Simmental bulls.Increasing the percentage of type Ⅰ muscle fiber and decreasing ⅡB type could improve beef tenderness.     

Relationships among glycolytic potential,dark cutting (dark,firm, and dry) beef,and cooked beef palatability

One hundred beef carcasses were selected at three packing plants and were used to determine the relationship between glycolytic potential (GP) and dark, firm, and dry (DFD) beef and to determine the effects of DFD status and GP on cooked beef palatability. Eight individual muscles were excised from one hindquarter of each carcass at d 7 postmortem: longissimus lumborum, psoas major, gluteus medius, tensor fasciae latae, rectus femoris, semimembranosus, biceps femoris, and semitendinosus. Ultimate pH, colorimeter readings, and Warner-Bratzler shear force were determined for all eight muscles at d 7 postmortem. A nine-member trained sensory panel evaluated cooked longissimus lumborum, gluteus medius, and semimembranosus steaks. Traits determined solely for the longissimus lumborum were GP (2 x [glycogen + glucose + glucose-6-phosphate] + lactate) and ether-extractable fat. A curvilinear relationship existed between GP and ultimate pH within the longissimus muscle. There appeared to be a GP threshold at approximately 100 micromol/g, below which lower GP was associated with higher ultimate pH and above which GP had no effect on ultimate pH. The greatest pH and muscle color differences between normal and DFD carcasses were observed in the longissimus lumborum, gluteus medius, semimembranosus, and semitendinosus muscles. Cooked longissimus from DFD carcasses had higher shear force values (46% greater) and more shear force variation (2.3 times greater variation) than those from normal carcasses. Dark cutting carcasses also had higher shear force values for gluteus medius (33% greater) and semimembranosus (36% greater) than normal carcasses. Sensory panel tenderness of longissimus, gluteus medius, and semimembranosus was lower for DFD carcasses than for normal carcasses. Longissimus and gluteus medius flavor desirability scores were lower for DFD than for normal carcasses. Steaks from DFD carcasses had more off-flavor comments than steaks from normal carcasses, specifically more "peanutty," "sour," and "bitter" flavors. The DFD effect of higher shear force values was approximately five times greater (+3.11 kg vs +0.63 kg) for carcasses with "slight" marbling scores than for carcasses with "small" marbling scores. In general, higher GP was associated with increased tenderness, even among normal carcasses. In conclusion, low GP was associated with DFD beef and resulted in substantially less-palatable cooked steaks.     

Recovering value from beef carcasses classified as dark cutters by United States Department of Agriculture graders

Effects of the dark-cutting condition were examined on commercially slaughtered beef carcass sides that were classified into groups exhibiting 1/3, 1/2, and full degrees of the dark-cutting (DEGDC) condition, as evaluated by a USDA-Agricultural Marketing Service grader (n = 20 per group). Twenty-nine muscles of each carcass side were evaluated to determine the ultimate pH and color (L*, a*, and b*). Fourteen beef muscles (biceps femoris, deep pectoral, chuck complexus, gluteus medius, infraspinatus, latissimus dorsi, psoas major, longissimus thoracis, longissimus lumborum, semimembranosus, semitendinosus, triceps brachii long head, tensor fasciae latae, and vastus lateralis) were evaluated using Warner-Bratzler Shear force (WBSF) and a trained sensory panel. The muscle x DEGDC interaction was significant for ultimate pH, L*, a*, and b* values (P < 0.05). When ultimate pH values of individual muscles were compared with the same muscles evaluated in a previous study, the 1/3, 1/2, and full DEGDC had 7, 9, and 5 muscles, respectively, that fell within a computed 95% prediction limit of what would be considered as a normal pH but were more variable as measured by within-class CV. Color values (L*, a*, and b*) of the muscles from dark-cutting carcasses were numerically lower than those from the normal carcasses. A survey designed to determine the ideal color range of beef lean for retail meat merchandisers (n = 34) and food service chefs (n = 33) across the United States resulted in data analyzed using principal components analysis of L*, a*, and b* values for muscles dissected in the study to estimate the true values for dark-cutting carcasses. Muscles that were within an acceptable color value range for food service chefs had the potential to add between $42.29 to $26.44 and $14.71 to $8.11 per side when valued at Choice and Select prices, respectively. Muscles that were within an acceptable color value range had the potential to add between $30.39 to $16.74 and $10.37 to $5.03 per side for retail meat merchandisers when acceptable muscles were valued at Choice and Select prices, respectively. No muscle x DEGDC interactions were detected for WBSF and sensory panel scores (P > 0.05), but differences were detected among muscles (P < 0.05). Several muscles were considered salvageable from the dark-cutting carcasses that were evaluated, and no significant differences in sensory scores or WBSF between DEGDC classes suggested equal sensory expectations for muscles from dark-cutting carcasses.     

Relationships between thyroid status, tissue oxidative metabolism, and muscle differentiation in bovine fetuses

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 (rT(3)) and thyroxine (T(4)) increased during development from day 110 to day 210 or 260, respectively, whereas concentration of triiodothyronine (T(3)) and hepatic type-1 5'-deiodinase activity (5'D1) increased from day 180 onwards. On day 260, high T(4) and rT(3) and low T(3) 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 T(4) and T(3)) 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 T(3) concentrations in the last gestational trimester could be involved in the differentiation of oxidative skeletal muscles.     

Muscle type‐specific effect of myostatin deficiency on myogenic regulatory factor expression in adult double‐muscled Japanese Shorthorn cattle

To clarify muscle type‐specific effect of myostatin on myogenic regulatory factors (MRFs), we examined mRNA expression of MRFs in five skeletal muscles of normal (NM) and myostatin‐deficient double‐muscled (DM) adult Japanese Shorthorn cattle by quantitative reverse‐transcribed PCR. Among the four MRFs, namely, Myf5, MyoD, myogenin, and MRF4, MyoD expression was different among the muscles of the DM cattle (P < 0.01) but not of the NM cattle. Meanwhile, MyoD expression was significantly elevated only in masseter (MS) muscle in the DM cattle due to the myostatin deficiency (P < 0.05). Myf5 and MRF4 expression in semitendinosus (ST) was higher in the DM than in the NM cattle (P < 0.05). According to analysis of myosin heavy chain (MyHC) isoform expression, more MyHC‐2x and ‐2a and less ‐slow isoforms were expressed in the longissimus and ST muscles compared to the MS muscle in both cattle (P < 0.05), but no significant difference in MyHC expression was observed between the NM and DM cattle. Taken together, myostatin has influences on Myf5 and MRF4 expression in faster‐type muscles and on MyoD expression in slower‐type muscles, suggesting a possible muscle type‐specific effect of myostatin in skeletal muscle growth and maintenance.     

Effects of postmortem aging and USDA quality grade on Warner-Bratzler shear force values of seventeen individual beef muscles

Forty USDA Select and 40 upper two-thirds USDA Choice beef carcasses were used to determine the effects of postmortem aging on tenderness of 17 individual beef muscles. Biceps femoris-long head, complexus, gluteus medius, infraspinatus, longissimus dorsi, psoas major, rectus femoris, semimembranosus, semitendinosus, serratus ventralis, spinalis dorsi, supraspinatus, tensor fasciae latae, teres major, triceps brachii-long head, vastus lateralis, and vastus medialis muscles were removed from each carcass. Seven steaks (2.54-cm thick) were cut from every muscle, and each steak was assigned to one of the following postmortem aging periods: 2, 4, 6, 10, 14, 21, or 28 d postmortem. After completion of the designated aging period, steaks were removed from storage (2 degrees C, never frozen), cooked to a peak internal temperature of 71 degrees C, and evaluated using Warner-Bratzler shear force (WBSF). Analysis of WBSF revealed a 3-way interaction (P = 0.004) among individual muscle, USDA quality grade, and postmortem aging period. With the exception of the Select teres major, WBSF of all muscles (both quality grades) decreased with increasing time of postmortem storage. Nonlinear regression was used to characterize the extent (aging response) and rate of decrease in WBSF from 2 through 28 d postmortem for each muscle within each quality grade. In general, WBSF of upper two-thirds Choice muscles decreased more rapidly from 2 to 10 d postmortem than did corresponding Select muscles. Muscles that had greater aging responses generally had greater 2-d WBSF values. The upper two-thirds Choice psoas major, serratus ventralis, and vastus lateralis muscles required similar aging times to complete a majority of the aging response (< or =0.1 kg of aging response remaining) compared with analogous Select muscles. The upper two-thirds Choice complexus, gluteus medius, semitendinosus, triceps brachii-long head, and vastus medialis muscles required 4 to 6 d less time to complete a majority of the aging response than did comparable Select muscles. Aging times for Select biceps femoris-long head, infraspinatus, longissimus dorsi, rectus femoris, semimembranosus, spinalis dorsi, supraspinatus, and tensor fasciae latae muscles were > or =7 d longer than those for corresponding upper two-thirds Choice muscles. Results from this study suggest that muscle-to-muscle tenderness differences depend on quality grade and aging time and that postmortem aging should be managed with respect to individual muscle and USDA quality grade.     

The extent of proteolysis is independent of sarcomere length in lamb longissimus and psoas major.

The objective of this experiment was to determine the effect of sarcomere length on postmortem proteolysis and meat tenderness. Eighteen Dorset market-weight sheep were slaughtered conventionally. The longissimus thoracis et lumborum and psoas major from each carcass were either left intact on the carcass (control), which was chilled at 0 degrees C, or excised from the carcass and chilled in an ice slurry (0 degrees C). At 24 h, control muscles were excised, and all muscles were cut into sections and assigned to 1 or 10 d of postmortem storage at 2 degrees C. Sarcomere length was shorter (P < .01), as intended, in the shortened relative to the control treatment and in longissimus relative to psoas major (1.36 vs 1.69 microm, raw longissimus; 1.45 vs 3.03 microm, raw psoas major). Sarcomere length was not affected (P > .05) by aging time. Western blot analysis of troponin-T and desmin indicated no effect (P > .05) of the shortened treatment compared to the control on the extent of proteolysis. Regardless of aging time or treatment, troponin-T was more degraded (P < .01) in longissimus than in psoas major (38.1 vs 23.5%) and desmin tended to be more degraded (P = .08) in longissimus than in psoas major (50.4 vs 35.1%). Regardless of muscle or treatment, aging 10 d compared to 1 d increased degradation of troponin-T (46.3 vs 15.3%) and desmin (69.3 vs 16.1%). Warner-Bratzler shear force was greater (P < .01) in the shortened treatment than in control (6.9 vs 3.8 kg), greater (P < .01) in longissimus than in the psoas major (6.5 vs 4.2 kg), and greater (P < .01) with 1 d than with 10 d of aging time (6.1 vs 4.6 kg). A muscle x aging time interaction (P < .05) indicated shear force declined more in longissimus than in psoas major during aging. We conclude that sarcomere length did not affect the extent of proteolysis. However, sarcomere length may have an indirect effect on tenderization during aging due to its effect on initial tenderness.     

Proteome analysis of whole and water‐soluble proteins in masseter and semitendinosus muscles of Holstein cows

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.     

Beef longissimus slice shear force measurement among steak locations and institutions

The objectives of this study were 1) to determine which longissimus thoracis et lumborum steaks were appropriate for slice shear force measurement and 2) to determine the among and within institution variation in LM slice shear force values of 6 institutions after they received expert training on the procedure and a standard kit of equipment. In experiment 1, longissimus thoracis et lumborum muscles were obtained from the left sides of 50 US Select carcasses. Thirteen longissimus thoracis and 12 longissimus lumborum steaks were cut 2.54 cm thick from each muscle. Slice shear force was measured on each steak. Mean slice shear force among steak locations (1 to 25) ranged from 19.7 to 27.3 kg. Repeatability of slice shear force (based on variance) among steak locations ranged from 0.71 to 0.96. In experiment 2, the longissimus thoracis et lumborum were obtained from the left sides of 154 US Select beef carcasses. Eight 2.54-cm-thick steaks were obtained from the caudal end of each frozen longissimus thoracis, and six 2.54-cm-thick steaks were obtained from the cranial end of each frozen longissimus lumborum. Seven pairs of consecutive steaks were assigned for measurement of slice shear force. Seven institutions were assigned to steak pairs within each carcass using a randomized complete block design, such that each institution was assigned to each steak pair 22 times. Repeatability estimates for slice shear force for the 7 institutions were 0.89, 0.83, 0.91, 0.90, 0.89, 0.76, and 0.89, respectively, for institutions 1 to 7. Mean slice shear force values were least (P <0.05) for institutions 3 (22.7 kg) and 7 (22.3 kg) and were greatest (P <0.05) for institutions 5 (27.3 kg) and 6 (27.6 kg). Institutions with greater mean slice shear force (institutions 5 and 6) used cooking methods that required more (P <0.05) time (32.0 and 36.9 min vs. 5.5 to 11.8 min) to reach the end point temperature (71 degrees C) and resulted in greater (P <0.05) cooking loss (both 26.6% vs. 14.4 to 24.1%). Differences among institutions in the repeatability of slice shear force were partially attributable to differences among institutions in the consistency of steak thawing and cooking procedures. These results emphasize the importance of sample location within the muscle and cooking method in the measurement of tenderness and indicate that with proper training and application of the protocol, slice shear force is a highly repeatable (R approximately 0.90) measure of beef LM tenderness.     

Increasing tenderness of beef round and sirloin muscles through prerigor skeletal separations

Crossbred steers (n = 30) were used to explore and compare tenderness improvements in beef round and sirloin muscles resulting from various methods of prerigor skeletal separations. Animals were slaughtered according to industry procedures, and at 60 min postmortem one of six treatments was applied to each side: A) control, B) saw pelvis at the sirloin-round junction, C) separate the pelvic-femur joint, D) saw femur at mid-point, E) combination of B and C, and F) combination of B and D. After 48 h, the following muscles were excised from each side: semimembranosus, biceps femoris, semitendinosus, and adductor from the round; vastus lateralis and rectus femoris from the knuckle; and gluteus medius, biceps femoris and psoas major from the sirloin. Following a 10-d aging period, samples were removed from each muscle to determine the effect of treatment on sarcomere length and Warner-Bratzler shear force. Most skeletal separation treatments resulted in longer sarcomeres than controls for semimembranosus, adductor, semitendinosus, and gluteus medius muscles. All skeletal separation treatments yielded shorter sarcomeres for the psoas major as compared with controls. Warner-Bratzler shear force differed among treatments for rectus femoris, semitendinosus, and psoas major. For rectus femoris, treatments C, D, E, and F resulted in lower (P < 0.05) shear values than for controls. Treatments B, D, and F increased shear force of the semitendinosus relative to controls (P < 0.05) within muscle. Treatment F resulted in higher shear force values for the PM than controls (P < 0.05). Correlations between sarcomere length and shear force were found to be low and quite variable among muscles. In general, treatments increased sarcomere length of several muscles from the sirloin/round region, but had mixed effects on shear force values.     

Intermuscular variation in tenderness: association with the ubiquitous and muscle-specific calpains

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.     

Contents of the glucose transporters GLUT1 and GLUT4 in oxidative and glycolytic muscles of goat kids and adult goats

The objective of this study was to elucidate, whether the impaired insulin sensitivity with regard to glucose utilisation in ruminating goats compared with suckling goat kids may be due to a reduced expression of the glucose transporters GLUT1 and GLUT4 in skeletal muscle. Muscle samples were removed from red, oxidative muscles (M. masseter, diaphragm) and white, glycolytic muscles (M. longissimus dorsi, and M. semitendinosus) of five goat kids fed with milk exchanger and nine adult goats of different stages of life. Samples were analysed for their GLUT1 and GLUT4 contents. The muscles were characterised metabolically by measuring the activities of isocitrate dehydrogenase (ICDH) and of lactate dehydrogenase. In all four analysed muscles the GLUT4 contents of adult goats were significantly (p < 0.05) lower than in goat kids. Significant differences concerning the GLUT4 contents in skeletal muscles were not detected in adult goats of different stages of life. The GLUT1 contents differed to a lower extend between goat kids and adult goats. The results of this investigation indicate that the impaired insulin sensitivity of adult compared with suckling ruminants is accompanied by or leads to a decreased GLUT4 expression.