calpastatin and Body-Weight

In this study, we characterized the effects of 25-hydroxyvitamin D3 (25-OH D3) and manipulated dietary cation-anion difference (DCAD) on the performance, urine pH, serum constituents, carcass traits, tissue residual vitamin D and its metabolites, beef tenderness, and mRNA and protein concentrations of Ca-dependent proteinases in LM using 24 cull native Korean cows. The cows were divided into 3 groups of 8: control, 25-OH D3 supplemented (25-OH D3), and manipulated DCAD plus 25-OH D3 supplemented (DCAD+25-OH D3). Cows receiving 25-OH D3 or DCAD+25-OH D3 were dosed with 125 mg of 25-OH D3 6 d before slaughter. The manipulated DCAD (-10 mEq/100 g of DM) diet was fed from 20 to 6 d (14 d) before slaughter. The DCAD+25-OH D3 treatment decreased urine pH and increased serum Ca concentrations. Although the vitamin D concentrations in LM, liver, and kidney were not affected by 25-OH D3 or DCAD+25-OH D3, muscle tissue 25-OH D3 concentrations were increased by both regimens. Serum 25-OH D3 concentrations were increased by 25-OH D3 supplementation, and the increase was even greater for DCAD+25-OH D3. The same pattern was observed for serum 1,25- (OH)2 D3. However, the LM concentration of 1,25-(OH)2 D3 was less for DCAD+25-OH D3 than for control. Although Ca concentrations of LM increased numerically in response to 25-OH D3 supplementation, no statistical differences in Warner-Bratzler shear force or sensory traits of LM were detected. The LM of cows receiving 25-OH D3 with or without manipulated DCAD had greater concentrations of mu-calpain and m-calpain mRNA, whereas the reverse was observed for calpastatin mRNA. Expression of mu-calpain protein was increased relative to control by DCAD+25-OH D3. The amount of 25-OH D3 and manipulated DCAD administered to cull native Korean cows was insufficient to improve tenderness of beef by increasing muscle Ca concentration. However, DCAD+25-OH D3 induced greater expressions of mu-calpain protein as well as mRNA.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Body Weight; Calcifediol; Calcitriol; Calcium-Binding Proteins; Calpain; Cattle; Diet; Gene Expression Regulation, Enzymologic; Korea; Male; Meat; Muscle, Skeletal

The effects of beef carcass weight on muscle pH/temperature profile and selected meat quality attributes were evaluated. Twenty-six carcasses from light (≤260kg, n=15) and heavy (≥290kg, n=11) feedlot steers were randomly allocated and stimulated with low voltage electrical stimulation (LVES) for 30s at 7min post-mortem (pm). Quality evaluations were carried out on samples from the Longissimus et lumborum (LL) muscle from the left side of each carcass. Heavier carcasses showed faster pH decline and slower (P<0.05) temperature decline at 45min, 3, 6, 12 and 24h pm. Heavier carcasses passed through the heat shortening window (i.e. at pH6, temperature was >35°C) but there was no sign of sarcomere shortening in any carcass. Significantly lower (P<0.05) shear force values were recorded in the heavier carcasses at 3days pm but at 14days pm, heavier carcasses had numerically lower but not significantly different shear force. Heavier carcasses produced numerically higher but not significant (P>0.05) drip loss at 3 and 14days pm as well as higher L* (meat lightness) (P<0.05) and C* (chroma) (P<0.05) values early (2days) pm. However, at 14days pm, there were no significant differences between the light and heavy carcasses in terms of L* and C*. No significant difference was observed between heavy and light carcasses in terms of H* at 2 and 14days pm. The study showed that heavier carcasses which favor slaughter house pricing can be produced and processed alongside lighter carcasses without significant detrimental effects on meat quality by using low voltage electrical stimulation (LVES).

Topics: Abattoirs; Animals; Body Composition; Body Weight; Calcium-Binding Proteins; Calpain; Cattle; Color; Electric Stimulation; Hydrogen-Ion Concentration; Meat-Packing Industry; Muscle, Skeletal; Postmortem Changes; Red Meat; Temperature; Time Factors

This investigation examined the mechanisms by which coronary perivascular adipose tissue (PVAT)-derived factors influence vasomotor tone and the PVAT proteome in lean versus obese swine.. Coronary arteries from Ossabaw swine were isolated for isometric tension studies. We found that coronary (P=0.03) and mesenteric (P=0.04) but not subcutaneous adipose tissue augmented coronary contractions to KCl (20 mmol/L). Inhibition of CaV1.2 channels with nifedipine (0.1 µmol/L) or diltiazem (10 µmol/L) abolished this effect. Coronary PVAT increased baseline tension and potentiated constriction of isolated arteries to prostaglandin F2α in proportion to the amount of PVAT present (0.1-1.0 g). These effects were elevated in tissues obtained from obese swine and were observed in intact and endothelium denuded arteries. Coronary PVAT also diminished H2O2-mediated vasodilation in lean and, to a lesser extent, in obese arteries. These effects were associated with alterations in the obese coronary PVAT proteome (detected 186 alterations) and elevated voltage-dependent increases in intracellular [Ca(2+)] in obese smooth muscle cells. Further studies revealed that the Rho-kinase inhibitor fasudil (1 µmol/L) significantly blunted artery contractions to KCl and PVAT in lean but not obese swine. Calpastatin (10 μmol/L) also augmented contractions to levels similar to that observed in the presence of PVAT.. Vascular effects of PVAT vary according to anatomic location and are influenced by an obese phenotype. Augmented contractile effects of obese coronary PVAT are related to alterations in the PVAT proteome (eg, calpastatin), Rho-dependent signaling, and the functional contribution of K(+) and CaV1.2 channels to smooth muscle tone.

Topics: Animals; Body Weight; Calcium-Binding Proteins; Coronary Artery Disease; Coronary Vessels; Cysteine Proteinase Inhibitors; Disease Models, Animal; Intra-Abdominal Fat; Isometric Contraction; Mesenteric Arteries; Muscle, Smooth, Vascular; Obesity; Proteomics; Subcutaneous Fat; Sus scrofa; Vasoconstriction

We examined the effects of trandolapril and candesartan on changes in the levels of sarcoglycans and dystrophin in the right ventricle of rats with the left coronary artery ligation. Hemodynamic and morphological alterations suggested the development of hypertrophy of the right ventricle and chronic heart failure by the 8th week. By the end of the 8th week, alpha- and beta-sarcoglycans and dystrophin were decreased. Increases in mu- and m-calpains in the hypertrophied right ventricle were associated with an elevation of casein-proteolytic activity in the cytosolic fraction. Oral administration of 3 mg/kg/day trandolapril or 1 mg/kg/day candesartan from the 2nd to 8th week after the left coronary artery ligation attenuated decreases in alpha-sarcoglycan and dystrophin and reduced the increased proteolytic activity. The results suggest that attenuation of decreases in sarcoglycans and dystrophin is a possible mechanism underlying trandolapril- and candesartan-mediated improvement of structural and functional alterations of the right ventricle in the coronary artery-ligated rat.

Topics: Angiotensin II Type 1 Receptor Blockers; Angiotensin-Converting Enzyme Inhibitors; Animals; Benzimidazoles; Biphenyl Compounds; Blood Pressure; Blotting, Western; Body Weight; Calcium-Binding Proteins; Calpain; Coronary Vessels; Cytosol; Dystrophin; Gene Expression; Heart Rate; Heart Ventricles; Indoles; Ligation; Male; Myocardial Infarction; Protein Isoforms; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sarcoglycans; Tetrazoles; Time Factors; Transcription, Genetic

Research was conducted to determine the effects of supplemental dietary vitamin D3 on DMI, carcass traits, Warner Bratzler shear (WBS) force, calpastatin activity, plasma minerals, pH (0, 3, 12, and 24 h after slaughter), water-holding capacity (WHC), and sensory characteristics of three muscles. Pre-slaughter vitamin D3 treatments included no supplemental vitamin D3, 6 x 106 IU (MIU) of vitamin D3 for 4 d, or 6 MIU of vitamin D3 for 6 d. Cattle were slaughtered and carcasses were chilled for 48 h before removal of steaks from the longissimus, gluteus medius, and biceps femoris muscles. Steaks were aged at 2 degrees C for 7, 14, or 21 d before cooking to a final internal temperature of 70 degrees C for WBS and sensory panel analysis. Dry matter intake was lower for steers supplemented with vitamin D3 for 4 or 6 d. Live and carcass weights were lower (P < 0.05) in steers supplemented with vitamin D3. Supplementing 6 MIU/6 d of vitamin D3 decreased (P < 0.05) WBS values of gluteus steaks (pooled over aging times). Longissimus steaks from steers supplemented with vitamin D3 for 6 d had lower (P < 0.05) WBS force values than these steaks from control steers or steers fed vitamin D3 for 4 d at 7 d postmortem. Biceps femoris steaks from steers receiving vitamin D3 for 4 d had higher WBS values than steaks from control steers at 14 and 21 d postmortem. Feeding vitamin D3 at 6 MIU for 6 d decreased (P < 0.05) the percentage of steaks that had WBS values > or = 3.86 kg for all steaks. Feeding vitamin D3 had no effect on palatability traits evaluated by trained panelists. Blood Ca concentrations were greater (P < 0.05) when vitamin D3 was fed and with increased vitamin D3 feeding time. Feeding vitamin D3 for 6 d (vs 4 d) delayed pH decline for all muscle types after 0, 3, and 12 h postmortem. Water-holding capacity was increased (P > 0.02) after 0 h, 24 h, and 21 d postmortem when vitamin D3 was fed and was greater at 0 and 24 h if vitamin D3 was fed for 6 d rather than 4 d. These data suggest that supplementing 6 MIU of vitamin D3 will decrease DMI and improve beef tenderness through increased blood plasma Ca concentrations and WHC.

Topics: Animals; Body Weight; Calcium; Calcium-Binding Proteins; Cattle; Cholecalciferol; Cysteine Proteinase Inhibitors; Dose-Response Relationship, Drug; Energy Intake; Food Handling; Male; Meat; Muscle, Skeletal; Postmortem Changes; Random Allocation; Taste; Time Factors

A study was conducted to examine the effects of bird age and muscle tissue type on calpain and calpastatin activities in turkey skeletal muscle. Enzymatic activities of calpains and calpastatin were found to vary with bird age and muscle type. Breast muscle from younger birds (age 5 wk) had higher mu-calpain, m-calpain, and calpastatin activities (P < 0.05) than breast muscle from older birds (9, 13, and 17 wk of age). Thigh muscle calpain activities were not affected by bird age, but thigh calpastatin activity was found to increase with age, with muscle from 17-wk-old birds having 35% higher activity than muscle from 13-wk-old birds. When extracted from 9-wk-old turkeys, breast muscle mu-calpain activity was 30% higher than thigh muscle mu-calpain. By 13 wk of age, breast muscle mu-calpain activity was 20% less than thigh mu-calpain. Thigh muscle m-calpain and calpastatin activities were found to be significantly higher (P < 0.05) than that found in breast muscle, with some values more than double in older birds (17 wk of age).

Topics: Aging; Animals; Body Weight; Calcium-Binding Proteins; Calpain; Cohort Studies; Hydrogen-Ion Concentration; Muscle, Skeletal; Time Factors; Turkeys

This experiment was conducted to investigate the effects of feeding a protein-free diet on mRNA levels of the calpain system in skeletal muscle of growing pigs during a 15-d feeding trial. Twenty crossbred barrows were divided into two dietary treatments: control or protein-free diet (mean initial weight for both groups: 38.3 kg). Daily diets were provided at 2.5 times energy for maintenance (twice a day). On d 0, 3, and 14, biopsies were taken from longissimus muscle between the third and fourth ribs (d 0 and 3) and between the fourth and fifth rib (d 14). On d 15, animals were slaughtered and longissimus muscles were dissected and analyzed for calpastatin, and mu- and m-calpain activity. From biopsies, mRNA level of skeletal muscle calpain, mu- and m-calpain, and calpastatin were measured using reversed transcription PCR. Subsequently, PCR products were quantified using ELISA. Feeding the protein-free diet lowered growth rate to almost zero. Only total level of mRNA of mu-calpain on d 14 was influenced by dietary treatments, being lower for the protein-free group than for the control group (P < .05). However, proteolytic activities were not different between treatments. Total RNA concentration in longissimus muscle decreased during the experiment for both treatments, but on d 14 this was more pronounced for the protein-free than for the control group (P < .05). If mRNA levels were corrected for this change, specific mRNA level on d 14 of skeletal muscle calpain and mu-calpain were lower (P < .05) for the protein-free than for the control group. These data suggest that activity of the components of the calpain system are differentially regulated.

Topics: Aging; Animals; Base Sequence; Biopsy; Body Weight; Calcium-Binding Proteins; Calpain; Diet; Diet, Protein-Restricted; DNA Primers; DNA Probes; DNA, Complementary; Eating; Enzyme-Linked Immunosorbent Assay; Male; Muscle, Skeletal; Polymerase Chain Reaction; RNA, Messenger; Swine; Weight Gain

The objective of this study was to determine the effects of dietary administration of a beta-adrenergic agonist (BAA; L-644,969) and male sex condition (ram vs wether) on muscle growth and meat quality of Dorset x Romonov lambs believed to be heterozygous for the callipyge gene. At approximately 17 wk of age, lambs were blocked by weight within each sex condition and randomly assigned to BAA treatment group. The interaction of BAA and male sex condition was not significant for any of the traits measured. Rams had greater initial and final live weights, average daily gain, and hot carcass weight (P < .01). Rams did not differ (P > .05) from wethers with respect to any of the carcass traits, possibly because the wethers were so lean and heavily-muscled that there was little room for improvement. Kidney-pelvic fat weight was reduced 26% by BAA (P < .05). Knife separable lean weight and whole carcass proximate composition were not affected (P > .05) by BAA or male sex condition. Administration of BAA increased calpastatin activity at 20 d (1.1 vs 1.5 units/g), but not at 0 h (3.9 vs 4.8 units/g) postmortem, decreased myofibril fragmentation index (60.7 vs 44.9), and increased shear force (8.2 vs 10.9 kg) at 20 d postmortem (P < .05). These data suggest that muscle growth rates are near maximum in lambs expressing the callipyge gene, regardless of male sex condition or BAA treatment. Therefore, it seems that the callipyge gene exerts most, but not all, of its effect through intracellular events similar to those initiated by administering BAA.

Topics: Adrenergic beta-Agonists; Animals; Body Composition; Body Weight; Calcium-Binding Proteins; Diet; Heterozygote; Hydrogen-Ion Concentration; Male; Meat; Muscle Development; Muscle, Skeletal; Orchiectomy; Pyridines; Random Allocation; Sheep; Temperature; Weight Gain

The theory that net muscle growth is, at least partly, regulated by catabolic factors has been tested in order to set up an animal model to study meat aging and post-mortem tenderization. Male and female chickens of a layer strain (White Leghorn), a commercial broiler strain (Ross), and two experimental broiler lines (designated GL and FC) were used to estimate differences in proteolytic enzyme activities in the breast muscles. The GL and the FC lines were selected for high body weight gain and high feed efficiency, respectively. At 6 wk of age the birds were slaughtered and the activities of endogenous proteinases and their specific inhibitors in breast muscles measured. The Leghorns showed significant differences in all traits compared with the three broiler genotypes. Within the broiler types, FC birds tended in the direction of the Leghorns and GL birds in the opposite direction. Ross birds were intermediate between FC and GL birds. All types and sexes differed significantly in slaughtering weight. Feed conversion ratio and protein conversion ratio were highest for Leghorns. The FC birds showed the lowest feed conversion. Ross and GL birds showed intermediate values. The Leghorns showed higher calpain activities and lower calpastatin activity than the three broiler genotypes. The FC broilers showed intermediate calpain and calpastatin activities but higher cathepsin H and total cystatin values. The GL broilers showed lower cathepsin B, D, and H activities. In all cases the Ross broilers showed intermediate values. From these figures it is concluded that the strains of birds used in this study can be used as a natural source of variability to study the mechanisms involved in post-mortem proteolytic degradation and thus in the study of muscle tenderization and meat aging. It is also concluded that it could be very interesting to study the behavior of the different proteolytic systems more carefully in relation to muscular growth characteristics and compare them to anabolic factors involved in muscle growth.

Topics: Animals; Body Weight; Calcium-Binding Proteins; Calpain; Cathepsins; Chickens; Female; Male; Muscle, Skeletal; Sex Factors; Species Specificity

The present experiment was conducted to determine the effect of the callipyge phenotype on traits affecting muscle growth and meat tenderness. Dorset wethers (N = 40) that were either carriers or non-carriers were fed grain and slaughtered at 169 d of age. Callipyge phenotype did not affect (P > .05) slaughter weight, hot carcass weight, or weights of the heart, spleen, viscera, kidney-pelvic fat, head, and pelt; however, callipyge lambs had a higher dressing percentage and lighter lungs, liver, and kidneys (P < .01). Callipyge lambs had reduced fat thickness and marbling score and higher leg scores and longissimus area (34%). Adductor (30%), biceps femoris (42%), gluteus group (31%), longissimus (32%), psoas group (20%), quadriceps femoris (18%), semimembranosus (38%), and semitendinosus (26%) weights were higher in the callipyge phenotype (P < .01); however, phenotype did not affect (P > .05) weights of infraspinatus or supraspinatus. Longissimus pH and temperature declines were not affected (P > .05) by phenotype. Longissimus myofibril fragmentation index was lower at 1 (27%), 7 (35%), and 21 (37%) d postmortem and Warner-Bratzler shear force was higher at 1, 7, and 21 d postmortem in the callipyge phenotype (P < .01). Shear force values of callipyge lambs at 21 d postmortem tended to be greater (P = .12) than shear force values of non-carriers at 1 d postmortem . Activities of calpastatin (83%) and m-calpain (45%) were higher in the callipyge (P < .01); however mu-calpain activity was not affected (P > .05). Longissimus and semitendinosus RNA concentration, DNA content, RNA content, protein content, and the RNA:DNA ratio were higher (P < .05), but DNA concentration, protein concentration, and protein:DNA were not affected in the callipyge phenotype. The higher calpastatin activity associated with callipyge suggests that protein degradation may be reduced in the live animal. Additionally, the increased muscle DNA content associated with the callipyge phenotype suggests an increase in satellite cell proliferation, and results in an increased capacity of skeletal muscle to accumulate and maintain myofibrillar protein. These results suggests that both reduced rate of protein degradation and higher capacity for protein synthesis are consequences of the callipyge condition.

Topics: Actinin; Animals; Blotting, Western; Body Weight; Calcium-Binding Proteins; Calpain; Connectin; Desmin; Hydrogen-Ion Concentration; Hypertrophy; Male; Meat; Muscle Development; Muscle Fibers, Skeletal; Muscle Proteins; Muscle, Skeletal; Nucleic Acids; Phenotype; Protein Kinases; Sheep; Sheep Diseases; Temperature; Troponin

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.

Topics: Animals; Body Weight; Calcium-Binding Proteins; Calpain; Chickens; Muscles; Substrate Specificity

Muscle proteolysis is controlled by a wide range of enzyme systems. The reported effects of the calcium dependent proteinases (calpain I and II) and its specific inhibitor (calpastatin) on myofibrillar structure, has led to the speculation that this system may have a pivotal role in regulating protein turnover and muscle growth. The present study highlights the possibility of protein degradation being subject to genetic variation. The relationship between genotype, level of nutrition, muscle protein turnover and the calpain system in young milk-fed lambs was assessed. Male lambs which had been selected for 10 generations for high (W+) and low (W-) weight at weaning were used in the study. Lambs were removed from their mothers 4 days after birth and surgically fitted with abomasal catheters and infused with reconstituted milk replacer at a high or a low rate. At 8 weeks of age, measurements of muscle protein gain, synthesis and degradation were performed, the animals were slaughtered and samples rapidly removed for subsequent chemical analysis. The liveweight gain and weight of the m vastus lateralis was reflected (P < 0.001) in the designed differences in nutrient supply. The weight of the m vastus lateralis was greater (P < 0.01) in the W+ compared to the W- lambs. The rate of protein synthesis and calculated degradation were greater (P < 0.05) in W+ than W- lambs. Calpain I and II and calpastatin activity were not significantly altered by genotype or nutrition. Calpastatin mRNA abundance increased significantly (P < 0.05) between 1 and 8 weeks of age. Regression analysis revealed genotype-specific responses with respect to calpastatin activity and mRNA abundance.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Analysis of Variance; Animal Nutritional Physiological Phenomena; Animals; Body Weight; Calcium-Binding Proteins; Cysteine Proteinase Inhibitors; Genetic Variation; Genotype; Male; Milk; Muscle Development; Muscle Proteins; Muscles; Phenylalanine; RNA, Messenger; Sheep

To investigate the role of calpains in myofibrillar protein degradation in skeletal muscle and the regulation of their activity in vivo, we studied the effects of fasting on gene expression of calpains and calpastatin in the skeletal muscle of rabbits. In response to fasting, myofibrillar protein degradation increased 2-fold and mRNA levels of calpain I, calpain II and calpastatin were also increased. However, calpain and calpastatin activities remained unchanged. To investigate this discrepancy, we analysed polysomal calpain mRNA. Results indicated that fasting caused a 2-fold increase in the loading of calpain I and II mRNAs on ribosomes. Thus transcription of genes encoding calpain may be increased during fasting to ensure adequate synthesis of the proteinases needed to mobilize muscle protein reserves. The effect of fasting on calpain and calpastatin mRNA expression is shared by cathepsin D and proteasome C2 but not by beta-actin, implying that fasting invokes control of several proteolytic systems in skeletal muscle and underscores the possibility that each proteolytic system plays a role in the adaptation of skeletal muscle to the fasted state.

Topics: Animals; Blotting, Western; Body Weight; Calcium-Binding Proteins; Calpain; Cathepsin D; Cysteine Endopeptidases; Fasting; Gene Expression; Multienzyme Complexes; Muscles; Polyribosomes; Proteasome Endopeptidase Complex; Rabbits; RNA, Messenger