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Biomedical subjects

F A Witzmann

Publications and source records attributed to F A Witzmann.

35 records · Page 2Linked to original sources

Soleus muscle atrophy in rats induced by cast immobilization: lack of effect by anabolic steroids.

To test the hypothesis that an anabolic steroid such as nandrolone decanoate (ND) will ameliorate or abolish disuse-mediated alterations resulting from five weeks of hindlimb immobilization, female Sprague-Dawley rats were divided into six groups: control, control + ND, shortened-immobilized, shortened-immobilized + ND, stretched-immobilized, and stretched-immobilized + ND. Immobilization was accomplished by wrapping hindlimbs with plaster of Paris. Nandrolone decanoate in sesame oil was administered via weekly intraperitoneal injection (7 mg/kg) while nontreated rats received equivalent volumes of the vehicle. After five weeks selected morphometric, biochemical, and mechanical parameters were examined in the slow-twitch soleus muscle (SOL). Muscle wet weight fell from a control value of 145 +/- 19 mg to 70 +/- 8 mg (p less than .05) in the immobilized-shortened group where ND had no effect (81 +/- 11 mg). Muscle stretch alone prevented weight loss (159 +/- 31 mg). The addition of ND resulted in significant SOL hypertrophy (200 +/- 42 mg), p less than .05. Immobilization resulted in a significant shift in muscle protein distribution toward sarcoplasmic protein, a change unaltered by ND but abolished by stretch. Muscle strength, as indicated by peak tetanic tension, fell 45% (p less than .05) as a result of shortened immobilization. Nandrolone decanoate had no effect on this condition, although the ameliorating effect of stretch was abolished when in combination with ND treatment. The elevation observed in maximal shortening velocity (shortened-immobilized) was unaffected by ND alone, but abolished by stretch with or without ND treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effect of ageing and exercise on skeletal muscle function.

The contractile and selected biochemical properties of fast- and slow-twitch skeletal muscle were studied at 9, 18, and 28 months of age in sedentary and regularly exercised rats. The isometric twitch duration was prolonged with aging in both the fast- and slow-twitch muscle. This effect was primarily due to a prolonged one-half relaxation time (1/2RT), which developed late in life. Regular exercise tended to further prolong the twitch duration, particularly in the slow-twitch soleus. Surprisingly, twitch and tetanic tension (Po), peak rate of tension development and decline, and the maximal shortening velocity were all unaltered between 9 and 28 months of age. Furthermore, regular exercise (running or swimming) had little or no effect on these properties. The prolonged 1/2RT with aging could not be explained by a decreased rate of Ca2+ sequestration by the sarcoplasmic reticulum, as the rate of Ca2+ uptake measured in muscle homogenates was unaltered in any of the muscles studied between 9 and 28 months. The degree of muscle fatigue (decline in Po) with 30 min of contractile activity in the slow-twitch soleus was not affected by aging. However, lactate reached two-fold higher levels and glycogen fell to considerably lower levels in the muscles of the old rats. This suggests an increased glycolysis and glycogen utilization during contractile activity in aged rats.

Adenosine Triphosphate↗

Effect of hindlimb immobilization on the fatigability of skeletal muscle.

The soleus (SOL) and extensor digitorum longus (EDL) muscles of the rat were studied in situ (33.5 degrees C) after 6 wk of disuse atrophy produced by hindlimb immobilization (IM). IM resulted in depressed peak twitch (Pt) and tetanic (Po) tension as well as a decreased rate of tension development (+dP/dt) and decline (-dP/dt) in the slow-twitch SOL. The fast-twitch EDL was affected to a lesser extent, because only Po (g . cm-2) and Po,-dP/dt (g . cm-2 . ms-1) were depressed after IM. Twitch duration, as measured by contraction time and one-half relaxation time, was shortened in the SOL and prolonged in the EDL. In both the fast and slow muscles 30 min of electrical stimulation resulted in a significant decline in Po. Relative to the prefatigued Po, the atrophied and control muscles showed a similar fatigue pattern. This occurred in spite of lower ATP and glycogen concentrations and higher lactate levels in the atrophied muscles. Our results indicate significant impairment of contractile function following IM in fast and slow muscles with preferential alterations in the slow SOL. In addition, despite a lower resting capacity and an increased dependence on glycolysis, contractile properties of atrophied muscles differed little from control muscles in their pattern of response to 30 min of electrical stimulation.

Adenosine Triphosphate↗

Acid phosphatase and protease activities in immobilized rat skeletal muscles.

The effect of hind-limb immobilization on selected lysosomal enzyme activities was studied in rat hind-limb muscles composed primarily of type I, IIA, or IIB fibers. Following immobilization, acid protease and acid phosphatase both exhibited significant (P less than 0.05) increases in their activity per unit weight in all three fiber types. Acid phosphatase activity increased at day 14 of immobilization in the three muscles and returned to control levels by day 21. Acid protease activity also changed biphasically, displaying a higher and earlier rise than acid phosphatase. The pattern of change in acid protease, but not acid phosphatase, closely parallels observed muscle wasting. The present data therefore demonstrate enhanced proteolytic capacity of all three fiber types early during muscular atrophy. In addition, the data suggest a dependence of basal hydrolytic and proteolytic activities and their adaptive response to immobilization on muscle fiber composition.

Acid Phosphatase↗

Muscle fatigue with prolonged exercise: contractile and biochemical alterations.

Alterations in the contractile and biochemical properties of fast and slow skeletal muscle were studied in rats following a prolonged swim to exhaustion. The exercise produced glycogen depletion (less than 1 mg/g tissue) in muscles representative of all three fiber types; the isometric contractile properties were altered in the 84% type I soleus (SOL) and the 60% type IIa extensor digitorium longus (EDL) but not in the 100% type IIb superficial region of the vastus lateralis (SVL). Peak tetanic tension (Po) and the rate of tension development and decline all decreased after prolonged exercise in both the SOL and the EDL. The maximal isotonic shortening velocity was highly correlated with the myofibrillar ATPase activity, and both were relatively resistant to fatigue. Furthermore, the Ca2+ sensitivity of the myofibrils was unaffected by exercise in both fast and slow muscle. The Ca2+ uptake capacity of the sarcoplasmic reticulum (SR) was reduced in both the SOL and the fast-twitch type IIa deep region of the vastus lateralis, whereas the SR ATPase activity was unchanged. Our findings provide evidence that prolonged exercise produces alterations in contractile and biochemical properties of type I and IIa but not type IIb fibers and that muscle fatigue as measured by a decline in Po is not necessarily correlated with glycogen depletion.

Adenosine Triphosphatases↗

Effect of thyrotoxicosis on sarcoplasmic reticulum in rat skeletal muscle.

The effect of thyrotoxicosis on the capacity of fragmented sarcoplasmic reticulum (FSR) and crude homogenate (CH) to sequester Ca2+ was determined in rat muscle for the slow-twitch type I soleus (SOL), the fast-twitch type IIA deep region of the vastus lateralis (DVL),and the fast-twitch type IIB superficial region of the vastus lateralis (SVL). The maximal rate of Ca2+ uptake (Vmax) and Km were determined in both the CH and FSR preparations, and the total Ca2+ uptake capacity of the CH was determined. In the slow SOL, thyrotoxicosis increased the Vmax (8.20 +/- 0.96 vs. 15.70 +/- 0.92 mumol Ca2+ . g wet muscle-1 . min-1) and the total Ca2+ uptake (17.62 +/- 1.30 vs. 27.13 +/- 2.16 mumol Ca2+ . g wet muscle-1) of the CH preparation. Thyrotoxicosis increased the FSR yield 2.3-fold in the slow-twitch SOL; however, the kinetic characteristics (Vmax and Km) of these vesicles were not altered. Thyrotoxicosis had no effect on the CH and FSR preparations in either the type IIA or type IIB sample. These results can be explained by a thyroid hormone-mediated increase in the quantity of the sarcoplasmic reticulum in type I muscle and suggest no effect on the hormone on the qualitative nature of the Ca2+-enzyme interaction.

Animals↗

Effect of disuse on sarcoplasmic reticulum in fast and slow skeletal muscle.

The effect of 6 wk of hindlimb immobilization on rat skeletal muscle sarcoplasmic reticulum (SR) was determined in the slow-twitch, type I soleus (SOL), the fast-twitch, type IIA deep region of the vastus lateralis (DVL), and the fast-twitch, type IIB superficial region of the vastus lateralis (SVL). Immobilization produced a significant decline in the Ca2+ uptake rate (Vmax) of SR vesicles from the slow SOL (0.930 +/- 0.116 to 0.365 +/- 0.071 mumol Ca2+ . mg-1 . min-1), while the SR Vmax increased in the fast SVL (2.763 +/- 0.133 to 5.209 +/- 0.687) and was unaltered in the DVL. Vesicles from the fast SVL and DVL also exhibited a higher total Ca2+ uptake capacity following immobilization. An evaluation of the time course of the immobilization-mediated effect revealed an increased Ca2+ uptake capacity in all three samples after 1 wk. In the SOL total Ca2+ uptake returned to control level after 2 wk, while in the fast-twitch muscles the higher capacities were maintained. The Ca2+-stimulated SR ATPase activity was not altered in any of the muscles studies, although the total SR ATPase activity increased twofold in the slow SOL.

Adenosine Triphosphatases↗

Recovery time course in contractile function of fast and slow skeletal muscle after hindlimb immobilization.

Contractile properties were evaluated in rats remobilized after 6 wk of hindlimb casting to evaluate the regenerative capacity of fast and slow skeletal muscles. Contractile parameters were determined in vitro (22 degrees C) in the type I soleus (SOL), type IIA and IIB extensor digitorum longus (EDL), and the type IIB superficial vastus lateralis (SVL). Immobilization (IM) shortened the SOL isometric twitch duration after which contraction time and half-relaxation time required 4 and 7 days to recover, respectively. In contrast, IM prolonged the twitch in the EDL and SVL and recovery required 14 and 7 days, respectively. Peak tetanic tension (g/cm2) fell in the SOL and EDL with IM and full recovery required 28 days. In this regard, the SVL remained unaltered. Rates of tension development and decline remained essentially unaltered in the fast muscles after IM but fell in the SOL, requiring 14 days to fully recover. Maximal shortening velocity, which had been elevated in all three muscles by IM, recovered rapidly. The present results demonstrate that both fast and slow muscle have the ability to completely recover from 6 weeks of IM.

Animals↗

Hindlimb immobilization: length-tension and contractile properties of skeletal muscle.

The effect of hindlimb immobilization (IM) on the contractile properties of fast and slow skeletal muscle was studied in rats following various periods of IM ranging from 1 to 42 days; muscle atrophy, muscle, fiber, and sarcomere length, and the length-tension characteristics were determined after 42 days of IM. The slow-twitch soleus (SOL), the fast-twitch extensor digitorum longus (EDL), and the fast-twitch superficial region of the vastus lateralis (SVL) all showed rapid atrophy following the onset of IM, reaching a new reduced steady-state weight by day 21. After 42 days of IM the passive tension (g) and active twitch tension (g/cm2) plotted vs. muscle length (cm) were shifted to the left for the slow-twitch SOL, indicating a decreased extensibility compared with control muscles. The peak tetanic tension of the slow SOL declined to 47% of the control level of 2,893 +/- 125 g/cm2, whereas the fast EDL maintained 72% of its initial force of 4,392 +/- 229 g/cm2, and the fast SVL was unaltered by IM. Peak twitch tension and peak rate of tension development and decline fell rapidly in the slow SOL while remaining relatively unaltered in the fast-twitch muscles. Surprisingly, maximal isotonic shortening velocity was elevated in both fast and slow muscles with IM. These results indicate that IM produces muscle atrophy in fast as well as slow skeletal muscle and, in addition, causes fiber type-specific changes in the contractile properties.

Animals↗

Adaptations in skeletal muscle following strength training.

Five men were studied before and after 7 wk of isokinetic strength training to determine its effects on muscle enzyme activities and fiber composition. One of the subject's legs was trained using 10 repeated 6-s maximal work bouts, while the other leg performed repeated 30-s maximal knee extension exercise. The total work accomplished by each leg was constant. Training 4 times/wk achieved similar gains in peak torque for both legs at the training velocity (3.14 rad/s) and at slower speeds. Fatigability of the knee extensor muscles, as measured by a 60-s exercise test, was similar in both legs after training. Biopsy specimens showed significant changes in the % of the muscle area composed of type I and IIa fibers as a result of both strength training programs. In terms of muscle enzymes, only the 30 s exercise program resulted in elevated glycolytic, ATP-CP and mitochondrial activities. Despite these changes, none of the parameters measured were found to be related to the gains in either muscle strength or fatigability during maximal isokinetic contractions.

Adaptation, Physiological↗

Lipid metabolism in skeletal muscle of endurance-trained males and females.

Bipsies were obtained from the gastrocnemius muscle of 13 male and 12 female distance runners and analyzed for [14C]palmitoyl-CoA oxidation, fiber composition, and the activities of selected enzymes. The male and female runners were similar in terms of maximal oxygen uptake (VO2max), training mileage, fiber compositions, and data collected during a 60-min treadmill run at 70% VO2max. Muscle succinate dehydrogenase and carnitine palmitoyl transferase activities were, however, significantly greater (P less than 0.05) in the male than in female runners. In addition, the male runners' muscle also showed a greater capacity to oxidize palmitoyl CoA. Little relationship, however, was found between muscle lipid metabolism, enzyme activities, and the calculated (respiratory exchange) fraction of energy derived from fat during 60 min of running at 70% VO2max. Although these data support the concept that endurance training (80-115 km/wk) markedly enhances the capacity of muscles to metabolize fats, the factors that regulate the usage of lipids during prolonged exercise do not appear to be limited by the capacity of the fibers to oxidize fatty acids, as determined by in vitro measurements.

Adult↗

Regional protein alterations in rat kidneys induced by lead exposure.

Lead is a potent neuro- and nephrotoxin in humans and a renal carcinogen in rats. Previous studies have detected lead-induced increases in the activities of specific detoxification enzymes in distinct kidney cell types preceding irreversible renal damage. While preferential susceptibility of the highly vascularized cortex to the effects of lead is clear, lead effects on the medullary region have remained unexplored. The present study was undertaken to investigate the extent to which regional renal protein expression differs and to determine which, if any, regionally distinct protein markers indicative of lead's renotoxic mechanism might be detected in kidney cortical and medullary cytosols. We examined protein expression in these two functionally and anatomically distinct regions, and identified several proteins that are differentially expressed in those regions and were significantly altered by lead. Kidney cytosols from rats injected with lead acetate (114 mg/kg, three consecutive daily injections) were separated by two-dimensional electrophoresis. Lead exposure significantly (P<0.001) altered the abundance (either or) of 76 proteins in the cortex and only 13 in the medulla. Eleven of the proteins altered in the protein patterns were conclusively identified either by matrix-assisted laser desorption/ionization mass spectrometry/electrospray ionization-mass spectrometry (MALDI-MS/ESI-MS) analysis of peptide digests, immunological methods, or by gel matching. Several of the cortical proteins altered by lead were unchanged in the medulla while others underwent similar but lesser alterations. These observations reflect the complexity of lead's nephrotoxicity and endorse the application of proteomics in mechanistic studies as well as biomarker development in a variety of toxicologic paradigms.

Amino Acid Sequence↗

Two-dimensional electrophoresis of precision-cut testis slices: toxicologic application.

Advances in tissue slice technology and a recent novel application of this technique to reproductive toxicology using bovine testis have demonstrated the remarkable utility of this approach. The objective of the present study was to combine this in vitro toxicity test system with large-scale two-dimensional polyacrylamide gel electrophoresis (2-DE) to detect and study alterations in testicular-slice protein patterns as molecular correlates of 1,3,5-trinitrobenzene (TNB) and 1,3-dinitrobenzene (DNB) toxicity. Previous studies have shown that testicular slices remain viable for > 24 h and, as measured by protein synthesis inhibition, TNB causes dose-related injury. Tissue-slices were prepared from bovine testicles incubated for 2, 4 or 6 h and exposed to either 100 microM, 500 microM or 1 mM DNB or TNB in the incubation medium. Slices were collected, solubilized, and separated by large scale 2-DE. Resulting protein patterns were then examined by image analysis, which revealed coefficients of variation in protein spot abundance comparable to patterns from fresh rodent tissue samples. Furthermore, specific protein alterations indicated dose-related inductions and declines in protein abundance, some progressive over time. The results of this investigation demonstrate the potential toxicologic utility of combining in vitro tissue-slice technology with high-resolution 2-DE protein mapping. The consolidation of these methods offers a novel approach for toxicity screening and testing, reduces experimental cost, and reduces the use of laboratory animals.

Animals↗