Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

Blanco, Cesar E.; Sieck, Gary C.; Edgerton, V. Reggie

doi:10.1007/bf01747468

Cited by 141 publications

(142 citation statements)

References 33 publications

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“…Sections were stained using laminin (Sigma) and N2.261 (developed by Dr. Helen M. Blau, obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA) antibodies to assess the fiber type-specific cross-sectional area and the percentage of myosin IIa isoforms and succinate dehydrogenase (SDH) activity to determine fiber type proportions (37,38). Digital images were obtained using an upright microscope with a camera (Axio Imager D1, Carl Zeiss, Wrek Göttingen, Germany), controlled by AxioVision AC software (AxioVision AC Rel.…”

Section: Methodsmentioning

confidence: 99%

Whole Body Deletion of AMP-activated Protein Kinase β2 Reduces Muscle AMPK Activity and Exercise Capacity

Steinberg

O’Neill

Dzamko

et al. 2010

Journal of Biological Chemistry

148

139

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AMP-activated protein kinase (AMPK) ␤ subunits (␤1 and ␤2) provide scaffolds for binding ␣ and ␥ subunits and contain a carbohydrate-binding module important for regulating enzyme activity. We generated C57Bl/6 mice with germline deletion of AMPK ␤2 (␤2 KO) and examined AMPK expression and activity, exercise capacity, metabolic control during muscle contractions, aminoimidazole carboxamide ribonucleotide (AICAR) sensitivity, and susceptibility to obesity-induced insulin resistance. We find that ␤2 KO mice are viable and breed normally. ␤2 KO mice had a reduction in skeletal muscle AMPK ␣1 and ␣2 expression despite up-regulation of the ␤1 isoform. Heart AMPK ␣2 expression was also reduced but this did not affect resting AMPK ␣1 or ␣2 activities. AMPK ␣1 and ␣2 activities were not changed in liver, fat, or hypothalamus. AICAR-stimulated glucose uptake but not fatty acid oxidation was impaired in ␤2 KO mice. During treadmill running ␤2 KO mice had reduced maximal and endurance exercise capacity, which was associated with lower muscle and heart AMPK activity and reduced levels of muscle and liver glycogen. Reductions in exercise capacity of ␤2 KO mice were not due to lower muscle mitochondrial content or defects in contraction-stimulated glucose uptake or fatty acid oxidation. When challenged with a high-fat diet ␤2 KO mice gained more weight and were more susceptible to the development of hyperinsulinemia and glucose intolerance. In summary these data show that deletion of AMPK ␤2 reduces AMPK activity in skeletal muscle resulting in impaired exercise capacity and the worsening of diet-induced obesity and glucose intolerance.The AMP-activated protein kinase (AMPK) 5 is an evolutionary conserved serine/threonine protein kinase that functions as a metabolic regulatory enzyme at both the intracellular and whole body level (1, 2). As a metabolic stress-sensing enzyme, AMPK is activated through phosphorylation of Thr 172 in the ␣-catalytic subunit by upstream kinases, liver kinase B1 (LKB1) and calcium/calmodulin-dependent kinase kinase in response to physiological processes that consume ATP (exercise) or inhibit ATP production (ischemia or hypoxia) (3). Following activation, AMPK acutely regulates lipid, protein, and carbohydrate metabolism through phosphorylation induced changes that alter enzyme activities by switching off ATP consuming anabolic pathways and switching on ATP generating catabolic pathways (4). In addition to these acute effects, AMPK regulates transcription factors to influence gene expression (4). Modulation of AMPK activity by hormones and cytokines adds a complex layer of regulation allowing energy supply and demand within a cell to be integrated with the energy requirements of the whole organism (5).AMPK functions as an ␣␤␥ heterotrimer where the C terminus of the ␤ isoforms (␤1 and ␤2) contains the subunit-binding sequence that is essential for binding the ␥ and ␣ subunits (6, 7). In addition to their structural role in maintaining the AMPK heterotrimer, AMPK ␤ subunits contain an evolutionary ...

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Section: Methodsmentioning

confidence: 99%

Whole Body Deletion of AMP-activated Protein Kinase β2 Reduces Muscle AMPK Activity and Exercise Capacity

Steinberg

O’Neill

Dzamko

et al. 2010

Journal of Biological Chemistry

148

139

View full text Add to dashboard Cite

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“…The reaction was terminated by multiple rinses in distilled water and then air-dried and placed under a coverslip. Individual muscle fibers were classified according to their SDH activity (high or low reactivity in EDL muscles; high or very high reactivity in soleus muscles) by interactive determination of the histochemical reaction intensity of individual fibers (3,14).…”

Section: Methodsmentioning

confidence: 99%

β₂-Agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol

Ryall

Gregorevic

Plant

et al. 2002

American Journal of Physiology-Regulatory, Integrative and Comp

100

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. ␤2-Agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol. Am J Physiol Regul Integr Comp Physiol 283: R1386-R1394, 2002. First published September 5, 2002 10.1152/ajpregu.00324.2002-Potential treatments for skeletal muscle wasting and weakness ideally possess both anabolic and ergogenic properties. Although the ␤2-adrenoceptor agonist clenbuterol has well-characterized effects on skeletal muscle, less is known about the therapeutic potential of the related ␤2-adrenoceptor agonist fenoterol. We administered an equimolar dose of either clenbuterol or fenoterol to rats for 4 wk to compare their effects on skeletal muscle and tested the hypothesis that fenoterol would produce more powerful anabolic and ergogenic effects. Clenbuterol treatment increased fiber cross-sectional area (CSA) by 6% and maximal isometric force (Po) by 20% in extensor digitorum longus (EDL) muscles, whereas fiber CSA in soleus muscles decreased by 3% and Po was unchanged, compared with untreated controls. In the EDL muscles, fenoterol treatment increased fiber CSA by 20% and increased Po by 12% above values achieved after clenbuterol treatment. Soleus muscles of fenoterol-treated rats exhibited a 13% increase in fiber CSA and a 17% increase in Po above that of clenbuteroltreated rats. These data indicate that fenoterol has greater effects on the functional properties of rat skeletal muscles than clenbuterol.

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“…The optimal pH of the preincubation solution was searched separately for each muscle and for each species in order to distinguish at least three levels of staining intensities after both acid and alkaline preincubations. Two additional serial sections were also stained for succinic dehydrogenase (SDH; Blanco et al 1988) and ␣ -glycerophosphate dehydrogenase (GPD; Martin et al 1985), and used to assess oxidative and glycolytic capabilities of single muscle fibers, respectively.…”

Section: Methodsmentioning

confidence: 99%

Evidence for Three Fast Myosin Heavy Chain Isoforms in Type II Skeletal Muscle Fibers in the Adult Llama (Lama glama)

Graziotti

Ríos

Rivero

2001

J Histochem Cytochem.

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(J-LLR)S U M M A R Y Skeletal muscle fiber types classified on the basis of their content of different myosin heavy chain (MHC) isoforms were analyzed in samples from hindlimb muscles of adult sedentary llamas ( Lama glama ) by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies, myofibrillar ATPase (mATPase) histochemistry, and quantitative histochemistry of fiber metabolic and size properties. The immunohistochemical technique allowed the separation of four pure (i.e., expressing a unique MHC isoform) muscle fiber types: one slow-twitch (Type I) and three fast-twitch (Type II) phenotypes. The same four major fiber types could be objectively discriminated with two serial sections stained for mATPase after acid (pH 4.5) and alkaline (pH 10.5) preincubations. The three fast-twitch fiber types were tentatively designated as IIA, IIX, and IIB on the basis of the homologies of their immunoreactivities, acid denaturation of their mATPase activity, size, and metabolic properties expressed at the cellular level with the corresponding isoforms of rat and horse muscles. Acid stability of their mATPase activity increased in the rank order IIA Ͼ IIX Ͼ IIB. The same was true for size and glycolytic capacity, whereas oxidative capacity decreased in the same rank order IIA Ͼ IIX Ͼ IIB. In addition to these four pure fibers (I, IIA, IIX, and IIB), four other fiber types with hybrid phenotypes containing two (I ϩ IIA, IIAX, and IIXB) or three (IIAXB) MHCs were immunohistochemically delineated. These frequent phenotypes (40% of the semitendinosus muscle fiber composition) had overlapped mATPase staining intensities with their corresponding pure fiber types, so they could not be delineated by mATPase histochemistry. Expression of the three fast adult MHC isoforms was spatially regulated around islets of Type I fibers, with concentric circles of fibers expressing MHC-IIA, then MHC-IIX, and peripherally MHC-IIB. This study demonstrates that three adult fast Type II MHC isoproteins are expressed in skeletal muscle fibers of the llama. The general assumption that the very fast MHC-IIB isoform is expressed only in small mammals can be rejected.

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Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

Cited by 141 publications

References 33 publications

Whole Body Deletion of AMP-activated Protein Kinase β2 Reduces Muscle AMPK Activity and Exercise Capacity

Whole Body Deletion of AMP-activated Protein Kinase β2 Reduces Muscle AMPK Activity and Exercise Capacity

β₂-Agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol

Evidence for Three Fast Myosin Heavy Chain Isoforms in Type II Skeletal Muscle Fibers in the Adult Llama (Lama glama)

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Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

Cited by 141 publications

References 33 publications

Whole Body Deletion of AMP-activated Protein Kinase β2 Reduces Muscle AMPK Activity and Exercise Capacity

Whole Body Deletion of AMP-activated Protein Kinase β2 Reduces Muscle AMPK Activity and Exercise Capacity

β2-Agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol

Evidence for Three Fast Myosin Heavy Chain Isoforms in Type II Skeletal Muscle Fibers in the Adult Llama (Lama glama)

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β₂-Agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol