scholarly journals CYTOCHEMICAL LOCALIZATION OF LACTIC DEHYDROGENASE IN WHITE SKELETAL MUSCLE

1964 ◽  
Vol 22 (1) ◽  
pp. 29-48 ◽  
Author(s):  
H. Dariush Fahimi ◽  
Chandra Raj Amarasingham
Keyword(s):  
Skeletal Muscle ◽  
White Muscle ◽  
Lactic Dehydrogenase ◽  
Gelatin Film ◽  
Histochemical Reaction ◽  
Cytochemical Localization ◽  
Tissue Sections ◽  
Modified Method ◽  
Muscle Extract ◽  
Adductor Magnus Muscle

The limitations of the conventional histochemical methods for localization of lactic dehydrogenase (LDH) in white skeletal muscle have been analyzed quantitatively. It is demonstrated that more than 80 per cent of LDH diffuses into the incubation medium within the first 10 minutes of incubation. Furthermore, it is confirmed that the addition of phenazine methosulfate (PMS) to the ingredients of the histochemical reaction for LDH increases substantially the capacity of the white muscle extract to reduce Nitro-BT. Based on these observations, a modified method of cytochemical localization of LDH has been developed. This method prevents the leakage of LDH from tissue sections by the application of all the ingredients of the histochemical reaction to tissue sections in a thin gelatin film. The incubation mixture contains PMS so that the staining system is independent of tissue diaphorase. The application of this method to the adductor magnus muscle of the rabbit revealed a fine reticulum in the sarcoplasm of all muscle fibers, in addition to the staining of mitochondria. The distribution of the staining suggests that LDH is localized in the sarcoplasmic reticulum.

Isozymes of lactic dehydrogenase in skeletal muscle

1962 ◽  
Vol 202 (5) ◽  
pp. 827-829 ◽  
Author(s):  
M. C. Blanchaer ◽  
M. Van Wijhe
Keyword(s):  
Skeletal Muscle ◽  
Guinea Pig ◽  
Dehydrogenase Activity ◽  
White Muscle ◽  
Lactic Dehydrogenase ◽  
Human Muscle ◽  
Isozyme Pattern ◽  
Rabbit Muscle ◽  
Red Muscle ◽  
Dehydrogenase Isozyme

The lactic dehydrogenase (LD) isozyme patterns of red and white muscle of the guinea pig, rabbit, and mouse were examined by electrophoresis in barbiturate-buffered agar. Red muscle consistently yielded five bands of lactic dehydrogenase activity, but white muscle usually produced only two bands corresponding to isozymes IV and V. Traces of isozyme III were noted occasionally in the white muscle of the rabbit and mouse, but bands I and II were absent from the white muscle of all three species. Crystalline rabbit muscle lactic dehydrogenase produced an isozyme pattern resembling that of a mixture of red and white rabbit muscle. The lactic dehydrogenase isozyme pattern of human muscle reported by previous workers resembles that of a mixture of the red and white muscle patterns.

Enzyme studies of skeletal muscle in mice with hereditary muscular dystrophy

1963 ◽  
Vol 205 (5) ◽  
pp. 897-901 ◽  
Author(s):  
Marilyn W. McCaman
Keyword(s):  
Skeletal Muscle ◽  
Muscular Dystrophy ◽  
Glutathione Reductase ◽  
Enzyme Activities ◽  
Lactic Dehydrogenase ◽  
Normal Muscle ◽  
Dystrophic Muscle ◽  
Nerve Section ◽  
Mouse Muscle ◽  
Dystrophic Mouse

The activities of 20 enzymes in normal, heterozygous, and dystrophic mouse muscle were studied by means of quantitative microchemical methods. Enzyme activities in normal and heterozygous muscle were essentially the same. In dystrophic muscle glucose-6-P dehydrogenase, 6-P-gluconic dehydrogenase, glutathione reductase, peptidase, ß-glucuronidase, and glucokinase activities were significantly higher than in normal muscle, while α-glycero-P dehydrogenase and lactic dehydrogenase activities were significantly lower. The pattern of enzyme activities found in normal gastrocnemius denervated by nerve section was strikingly similar to that in dystrophic muscle.

CHANGES IN THE RESTING POTENTIAL OF SKELETAL MUSCLE IN RATS WITH COLD ACCLIMATION

1966 ◽  
Vol 44 (5) ◽  
pp. 791-802 ◽  
Author(s):  
M. H. Sherebrin ◽  
A. C. Burton
Keyword(s):  
Skeletal Muscle ◽  
Electrical Properties ◽  
Cold Acclimation ◽  
White Muscle ◽  
Single Cells ◽  
Temperature Gradients ◽  
Resting Potential ◽  
The Mean ◽  
Shivering Thermogenesis ◽  
Main Factor

The resting potential of single cells in the flexor thigh muscles of rats was measured in an attempt to find a change in the electrical properties of the cell membrane with cold acclimation, in order to identify and relate metabolic changes occurring with non-shivering thermogenesis. The mean resting potential of cells in cold-acclimated rats was found to be slightly but significantly higher than in the controls. A larger temperature gradient with depth was measured in the cold-acclimated animals than in the controls. If the Q10 of resting potential with temperature is as great as 1.16, the higher potential in the cold-acclimated rats may be accounted for by this temperature difference. The resting potential was also found to vary with depth in both groups of rats. This could not be attributed to temperature gradients, and change from red to white muscle cells with depth is thought to be the main factor for the increase of potential with depth.

scholarly journals Low intrinsic running capacity is associated with reduced skeletal muscle substrate oxidation and lower mitochondrial content in white skeletal muscle

2011 ◽  
Vol 300 (4) ◽  
pp. R835-R843 ◽  
Author(s):  
Donato A. Rivas ◽  
Sarah J. Lessard ◽  
Misato Saito ◽  
Anna M. Friedhuber ◽  
Lauren G. Koch ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Insulin Sensitivity ◽  
Aerobic Capacity ◽  
Artificial Selection ◽  
White Muscle ◽  
Metabolic Diseases ◽  
Metabolic Health ◽  
Mitochondrial Content ◽  
Red Muscle ◽  
Selection For

Chronic metabolic diseases develop from the complex interaction of environmental and genetic factors, although the extent to which each contributes to these disorders is unknown. Here, we test the hypothesis that artificial selection for low intrinsic aerobic running capacity is associated with reduced skeletal muscle metabolism and impaired metabolic health. Rat models for low- (LCR) and high- (HCR) intrinsic running capacity were derived from genetically heterogeneous N:NIH stock for 20 generations. Artificial selection produced a 530% difference in running capacity between LCR/HCR, which was associated with significant functional differences in glucose and lipid handling by skeletal muscle, as assessed by hindlimb perfusion. LCR had reduced rates of skeletal muscle glucose uptake (∼30%; P = 0.04), glucose oxidation (∼50%; P = 0.04), and lipid oxidation (∼40%; P = 0.02). Artificial selection for low aerobic capacity was also linked with reduced molecular signaling, decreased muscle glycogen, and triglyceride storage, and a lower mitochondrial content in skeletal muscle, with the most profound changes to these parameters evident in white rather than red muscle. We show that a low intrinsic aerobic running capacity confers reduced insulin sensitivity in skeletal muscle and is associated with impaired markers of metabolic health compared with high intrinsic running capacity. Furthermore, selection for high running capacity, in the absence of exercise training, endows increased skeletal muscle insulin sensitivity and oxidative capacity in specifically white muscle rather than red muscle. These data provide evidence that differences in white muscle may have a role in the divergent aerobic capacity observed in this generation of LCR/HCR.

Influence of acclimation temperature on mitochondrial DNA, RNA, and enzymes in skeletal muscle

1998 ◽  
Vol 275 (3) ◽  
pp. R905-R912 ◽  
Author(s):  
Brendan James Battersby ◽  
Christopher D. Moyes
Keyword(s):  
Skeletal Muscle ◽  
Mitochondrial Dna ◽  
Steady State ◽  
Muscle Fiber ◽  
Copy Number ◽  
White Muscle ◽  
Fiber Types ◽  
Mitochondrial Enzymes ◽  
Dna Copy Number ◽  
Mitochondrial Content

Skeletal muscle fibers typically undergo modifications in their mitochondrial content, concomitant with alterations in oxidative metabolism that occur during the development of muscle fiber and in response to physiological stimuli. We examined how cold acclimation affects the mitochondrial properties of two fish skeletal muscle fiber types and how the regulators of mitochondrial content differed between tissues. After 2 mo of acclimation to either 4 or 18°C, mitochondrial enzyme activities in both red and white muscle were higher in cold-acclimated fish. No significant differences were detected between acclimation temperatures in the abundance of steady-state mitochondrial mRNA (cytochrome- c oxidase 1, subunit 6 of F0F1-ATPase), rRNA (16S), or DNA copy number. Steady-state mRNA for nuclear-encoded respiratory (adenine nucleotide translocase 1) and glycolytic genes showed high interindividual variability, particularly in the cold-acclimated fish. Although mitochondrial enzymes were 10-fold different between the two muscle types, mitochondrial DNA copy number differed only 4-fold. The relative abundance of mitochondrial mRNA and nuclear mRNA in red and white muscle reflected the differences in copy number of their respective genes. These data suggest that the response to physiological stimuli and determination of tissue-specific mitochondrial properties likely result from the regulation of nuclear-encoded genes.

EFFECTS OF FEEDING AND FOOD DEPRIVATION ON OXYGEN CONSUMPTION, MUSCLE PROTEIN CONCENTRATION AND ACTIVITIES OF ENERGY METABOLISM ENZYMES IN MUSCLE AND BRAIN OF SHALLOW-LIVING (SCORPAENA GUTTATA) AND DEEP-LIVING (SEBASTOLOBUS ALASCANUS) SCORPAENID FISHES

1993 ◽  
Vol 181 (1) ◽  
pp. 213-232 ◽  
Author(s):  
T. H. Yang ◽  
G. N. Somero
Keyword(s):  
Skeletal Muscle ◽  
Metabolic Rate ◽  
Food Deprivation ◽  
Enzyme Activities ◽  
Protein Concentration ◽  
White Muscle ◽  
Muscle Protein ◽  
Metabolism Enzymes ◽  
Red Muscle ◽  
Ad Libitum

The effects of feeding and fasting were examined on the deep-living short-spine thornyhead (Sebastolobus alascanus) and the confamilial shallow-living spotted scorpionfish (Scorpaena guttata) to determine whether the low metabolic rate of the deeper-living species was in part a consequence of food deprivation in its habitat. Laboratory acclimation for periods of 90–115 days under either ad libitum feeding or complete fasting did not lead to similar rates of respiration in individuals of the two species held under identical conditions. Respiration of fish fed ad libitum was 52 % (S. guttata) or 68 % (S. alascanus) higher than for fasted fish of the same species. Furthermore, the metabolic rates of freshly collected specimens of S. alascanus resembled those of laboratory-fasted fish. In white skeletal muscle, both total protein concentration and the activities of four enzymes of ATP metabolism, lactate dehydrogenase (LDH) and pyruvate kinase (PK) of glycolysis, malate dehydrogenase (MDH) and citrate synthase (CS, a citric acid cycle indicator), were lower in S. alascanus than in S. guttata. Within a species, protein concentration and activities of the four enzymes in white muscle, but not in brain, were higher in fed than in starved fish, although these differences were greater in S. alascanus than in S. guttata. During fasting, LDH and PK activity in white muscle of S. alascanus decreased much more than MDH and CS activity; decreases in enzyme activities in red muscle were smaller than those in white muscle. Activities of enzymes in white skeletal muscle of field-collected S. alascanus generally resembled those of the fasted specimens. In contrast, red muscle of field- collected S. alascanus, compared with that of either fed or starved laboratory-held specimens, had a highly glycolytic poise (high LDH and PK activities relative to MDH and CS activities), which may suggest that muscle enzyme activities in the field-collected fish reflect adaptation to the low oxygen level in its adult habitat, the oxygen minimum layer. The strong correlations found between tissue biochemical properties and respiration rate allow us to develop a predictive index for metabolic rate from simple biochemical analyses, e.g. white muscle protein content or CS activity. We conclude that the low metabolic rate of S. alascanus is due to at least four depth-related factors: reduced abundance of food, low temperature, low ambient oxygen concentration and darkness, which may select for reduced locomotory activity.

scholarly journals Sea Cucumber Peptides Improved the Mitochondrial Capacity of Mice: A Potential Mechanism to Enhance Gluconeogenesis and Fat Catabolism during Exercise for Improved Antifatigue Property

2020 ◽  
Vol 2020 ◽  
pp. 1-17
Author(s):  
Yihao Yu ◽  
Guoqing Wu ◽  
Yuge Jiang ◽  
Bowen Li ◽  
Chuanxing Feng ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Antioxidant Capacity ◽  
Sea Cucumber ◽  
Lactic Dehydrogenase ◽  
Mrna Levels ◽  
Energy Regulation ◽  
Lipid Catabolism ◽  
Swimming Test ◽  
Grip Test ◽  
Mitochondrial Capacity

Sea cucumber promotes multifaceted health benefits. However, the mechanisms of sea cucumber peptides (Scp) regulating the antifatigue capacity is still unknown. The present study is aimed at further elucidating the effects and mechanisms of Scp on the antifatigue capacity of mice. At first, C57BL/6J mice were assigned into four groups named Con, L-Scp, M-Scp, and H-Scp and received diets containing Scp (0%, 0.15%, 0.3%, and 0.5%, respectively) for continuous 30 days. On the 21th day, a fore grip test was conducted on mice. On the 25th day, a rotating rod test was conducted on mice. On the 30th day, the quantities of glycogen and mitochondrial DNA (mtDNA) were determined in 8 random mice and another 8 mice were forced to swim for 1 hour before slaughter for detecting biochemical indicators. It was observed that the Scp groups significantly prolonged the running time in rotarod, increased forelimb grip strength, improved lactic acid (LD) and urea nitrogen (BUN) levels in the serum, decreased lactic dehydrogenase (LDH) and glutamic oxalacetic transaminase (GOT) activities in the serum, increased blood glucose (BG) and glycogen (GN) levels in the liver and skeletal muscle after swimming, increased the activity of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in the skeletal muscle and heart, and improved antioxidant capacity. Furthermore, Scp treatment significantly elevated the mRNA and protein relative levels of power-sensitive factors, lipid catabolism, and mitochondrial biogenesis and significantly upregulated mRNA levels of gluconeogenesis. Besides, mtDNA before the swimming test was increased in the three Scp groups. These results show that Scp treatment has antifatigue capacity. Furthermore, these results suggest that improved energy regulation and antioxidant capacity may be the result of improved mitochondrial function.

Cardiac and skeletal muscle mitochondria have a monocarboxylate transporter MCT1

1999 ◽  
Vol 87 (5) ◽  
pp. 1713-1718 ◽  
Author(s):  
George A. Brooks ◽  
Marcia A. Brown ◽  
C. E. Butz ◽  
James P. Sicurello ◽  
Hervé Dubouchaud
Keyword(s):  
Skeletal Muscle ◽  
Striated Muscle ◽  
Lactic Dehydrogenase ◽  
Monocarboxylate Transporter ◽  
Lactate Oxidation ◽  
Western Blots ◽  
Lactate Shuttle ◽  
Muscle Mitochondria ◽  
Monocarboxylate Transporter 1 ◽  
Skeletal Muscle Mitochondria

To evaluate the potential role of monocarboxylate transporter-1 (MCT1) in tissue lactate oxidation, isolated rat subsarcolemmal and interfibrillar cardiac and skeletal muscle mitochondria were probed with an antibody to MCT1. Western blots indicated presence of MCT1 in sarcolemmal membranes and in subsarcolemmal and interfibrillar mitochondria. Minimal cross-contamination of mitochondria by cell membrane fragments was verified by probing for the sarcolemmal protein GLUT-1. In agreement, immunolabeling and electron microscopy showed mitochondrial MCT1 in situ. Along with lactic dehydrogenase, the presence of MCT1 in striated muscle mitochondria permits mitochondrial lactate oxidation and facilitates function of the “intracellular lactate shuttle.”

scholarly journals AN OSMIOPHILIC DISTYRYL DITETRAZOLIUM SALT (DS-NBT) FOR ULTRASTRUCTURAL DEHYDROGENASE ACTIVITY

1971 ◽  
Vol 19 (5) ◽  
pp. 273-285 ◽  
Author(s):  
ARNOLD M. SELIGMAN ◽  
IZHAK NIR ◽  
ROBERT E. PLAPINGER
Keyword(s):  
Skeletal Muscle ◽  
Dehydrogenase Activity ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Structural Features ◽  
Rat Skeletal Muscle ◽  
Succinic Dehydrogenase Activity ◽  
Tissue Penetration ◽  
Diaphorase Activity ◽  
Doubly Charged

The preparation of a new osmiophilic ditetrazolium salt (DS-NBT) containing two styryl groups, but retaining the essential structural features of nitro-BT, is given. DS-NBT may be used to demonstrate succinic dehydrogenase activity (SDH) faster than with nitro-BT and fully as rapidly as with TC-NBT. DS-NBT is more stable than TC-NBT and it is more reliably and more readily osmicated than TC-NBT. The ultrastructural sites of SDH and NADH2 diaphorase activity are similar to those shown for cytochrome oxidase activity with diaminobenzidine in rat heart. The sites of lactic dehydrogenase activity are found in the sarcoplasmic reticulum and mitochondria of rat skeletal muscle fixed in formaldehyde for 3 hr. Tissue penetration of the doubly charged ditetrazolium salt (DS-NBT) was facilitated by 40-sec sonication and by 5% dimethyl sulfoxide. Illustrative examples and controls are included.

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