Quantitative histochemical determination of muscle enzymes: biochemical verification.

Martin, Tom; Vailas, A. C.; Durivage, James B.; Edgerton, V. Reggie; Castleman, Kenneth R.

doi:10.1177/33.10.4045183

Cited by 100 publications

(69 citation statements)

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“…13 Quantitative histochemical determination of succinic dehydrogenase activity (SDH) and a-glycerol phosphate dehydrogenase activity (GPDH) were used as estimates of oxidative and glycolytic energy supply, respectively, essentially as done previously. 9,10 SDH and GPDH activities were determined by the microdensitometric technique described by Blanco et al, 14 and Martin et al, 15 respectively. Images were captured using the same tools as for ®ber typing with the exception that a narrow pass interference ®lter with peak emission of 570 nm was used so as to assess maximal absorption of Nitro-blue tetrazolium-diformazan, the in-vitro reaction end product.…”

Section: Histochemical Analysesmentioning

confidence: 99%

Effects of testosterone replacement therapy on skeletal muscle after spinal cord injury

Gregory

Vandenborne

Huang³

et al. 2002

Spinal Cord

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Study design: Randomized control. Objective: To examine the e ects of testosterone replacement therapy (TRT) on skeletal muscle 11 weeks after complete SCI. Setting: Athens, Georgia USA. Methods: Soleus (SOL), gastrocnemius (GA), tibialis anterior (TA), vastus lateralis (VL) and triceps brachii (TRI) muscles were taken from twelve young male Charles River rats 11 weeks after complete SCI (T-9 transection, n=8) or sham surgery (n=4). Rats received either TRT (two 5 cm capsules, n=4) or empty capsules (n=8) implanted at surgery. Muscle samples were sectioned and ®bers analyzed qualitatively for myosin ATPase and quantitatively for succinate dehydrogenase (SDH), a-glycerol-phosphate dehydrogenase (GPDH) and actomyosin ATPase (qATPase) activities using standard techniques. Results: SCI decreased average ®ber size (49+4%) in a ected muscles and the percentage of slow ®bers in SOL (93+3% to 17+2%). In addition, there was a decrease in SDH and an increase in GPDH and qATPase activities across the four hind-limb muscles of the SCI animals. Fiber size in the TRI was increased (31+2%) by SCI while enzyme activities were not altered. Average ®ber size across the four hind limb muscles was decreased by only 30% in TRT SCI animals and their SOL contained 39+2% slow ®bers. TRT also attenuated changes in enzyme activities. There was no e ect of TRT on the TRI relative to SCI.

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Section: Histochemical Analysesmentioning

confidence: 99%

Effects of testosterone replacement therapy on skeletal muscle after spinal cord injury

Gregory

Vandenborne

Huang³

et al. 2002

Spinal Cord

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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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“…For COX staining, the sections were incubated in the solution composed of 2 mg/ml of catalase (Sigma-Aldrich), 1 mg/ml of cytochrome c (Sigma-Aldrich), and 0.5 mg/ml of diaminobenzidine (Wako) in 0.1 M of phosphate buffer (pH 7.4) for 1 hour at 37°C. The SDH staining was performed according to the modified method of Martin et al 53 In brief, we prepared nitro blue tetrazolium stock composed of KCN 6.5 mg (Nacalai, Kyoto, Japan), EDTA 185 mg (Sigma-Aldrich), and nitro blue tetrazolium 100 mg (Nacalai) in 100 ml of 0.1 M phosphate buffer (pH 7.6) and 500 mM of sodium succinate solution (Nacalai) diluted in distilled water. Then the sections were incubated in a solution mixed with 2 ml of nitro blue tetrazolium stock solution, 0.2 ml of succinate solution, and 0.7 mg of phenazine methosulfate (Nacalai) for 15 minutes at 37°C.…”

Section: Cox/sdh Stainingmentioning

confidence: 99%

High-Fat Diet–Induced Lysosomal Dysfunction and Impaired Autophagic Flux Contribute to Lipotoxicity in the Kidney

Yamamoto¹,

Takabatake²,

Takahashi³

et al. 2016

JASN

204

194

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Excessive fat intake contributes to the progression of metabolic diseases via cellular injury and inflammation, a process termed lipotoxicity. Here, we investigated the role of lysosomal dysfunction and impaired autophagic flux in the pathogenesis of lipotoxicity in the kidney. In mice, a high-fat diet (HFD) resulted in an accumulation of phospholipids in enlarged lysosomes within kidney proximal tubular cells (PTCs). In isolated PTCs treated with palmitic acid, autophagic degradation activity progressively stagnated in association with impaired lysosomal acidification and excessive lipid accumulation. Pulse-chase experiments revealed that the accumulated lipids originated from cellular membranes. In mice with induced PTC-specific ablation of autophagy, PTCs of HFD-mice exhibited greater accumulation of ubiquitin-positive protein aggregates normally removed by autophagy than did PTCs of mice fed a normal diet. Furthermore, HFD-mice had no capacity to augment autophagic activity upon another pathologic stress. Autophagy ablation also exaggerated HFD-induced mitochondrial dysfunction and inflammasome activation. Moreover, renal ischemia-reperfusion induced greater injury in HFD-mice than in mice fed a normal diet, and ablation of autophagy further exacerbated this effect. Finally, we detected similarly enhanced phospholipid accumulation in enlarged lysosomes and impaired autophagic flux in the kidneys of obese patients compared with nonobese patients. These findings provide key insights regarding the pathophysiology of lipotoxicity in the kidney and clues to a novel treatment for obesity-related kidney diseases.

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Quantitative histochemical determination of muscle enzymes: biochemical verification.

Cited by 100 publications

References 0 publications

Effects of testosterone replacement therapy on skeletal muscle after spinal cord injury

Effects of testosterone replacement therapy on skeletal muscle after spinal cord injury

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

High-Fat Diet–Induced Lysosomal Dysfunction and Impaired Autophagic Flux Contribute to Lipotoxicity in the Kidney

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