Enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier

Noorden, C. J. F. Van; Bhattacharya, R. D.; Vogels, I. M. C.

doi:10.1016/s0065-1281(83)80077-4

Cited by 17 publications

(2 citation statements)

References 20 publications

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“…SDH is a membrane-bound enzyme located on the inner membrane of mitochondria (Lehninger, 1975). Van Noorden et al (1983) demonstrated that in the histochemical reaction for SDH using mPMS and azide, the reaction indicator (tetranitroblue tetrazolium formazan) was deposited in an intracellular site that corresponds to the location of mitochondria in rat hepatocytes. In muscle fibres, the mitochondria are located throughout the central core, alongside the myofibrils, and in the subsarcolemmal region (Eisenberg et al, 1974;Eisenberg & Kuda, 1975, 1976, 1977.…”

Section: Discussionmentioning

confidence: 99%

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

1988

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A histochemical technique was developed for the quantitative determination of succinic dehydrogenase (SDH) activity in muscle cross-sections using 1-methoxyphenazine methosulphate (mPMS) as the exogenous electron carrier, and azide as an inhibitor of cytochrome oxidase. The optimal composition of the incubation medium for the SDH reaction was determined. This histochemical procedure was compared to one using phenazine methosulphate (PMS) instead of mPMS and cyanide instead of azide. The substitution of mPMS and azide resulted in a substantial decrease in the non-specific reduction of nitroblue tetrazolium (NBT; the reaction indicator), i.e., 'nothing dehydrogenase' activity. With mPMS and azide in the reaction medium, the production of NBT formazan was linear for at least 9 min during the enzymic reaction. This compared to a non-linear reduction of NBT during the initial stages of the reactions (SDH and 'nothing dehydrogenase') when using PMS and cyanide. The use of both mPMS and azide also eliminated the production of NBT monoformazan which occurred with PMS and cyanide. This procedure was shown to meet various criteria established for the quantification of histochemical reactions.

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Section: Discussionmentioning

confidence: 99%

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

1988

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“…The embedded spinal cord was cooled on ice, placed on a cryostat chuck, and frozen in Freon at -150~ Serial transverse sections were cut (10 blm) in a cryostat cabinet at -25~ and mounted on slides coated with 3-amino-propyltriethoxysilane, dried at room termperature for ten minutes, and stored in liquid nitrogen. They were brought to room temperature for ten minutes, and stored in liquid nitrogen, prior to the histochemical treatment and reacted for one of the following enzymes, according to protocols previously described: gtucose-6-phosphate dehydrogenase (G6PDH; Van Noorden, 1984) succinate dehydrogenase (SDH; Van Noorden et al, 1983), and NADH tetrazolium reductase (NADH-TR; Lojda et al, 1976, and as modified by Van Raamsdonk et al, 1987). The sections were incubated for 20 min at 25~ for G6PDH and for 20 min at 37~ for both SDH and NADH-TR.…”

Section: Enzyme Histochemistrymentioning

confidence: 99%

Changes in enzyme histochemical profiles of identified spinal motoneurons of the European eel,Anguilla anguilla, following cordotomy

et al. 1996

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The enzyme histochemical profiles of glucose-6-phosphate dehydrogenase (a marker of synthetic performance), succinate dehydrogenase (an indicator of oxidative metabolism), and NADH tetrazolium reductase (a marker of overall neuronal activity) were determined for identified white muscle motoneurons in six control and six cordotomized cels. Images were digitized and mean integrated absorbances obtained using appropriate hardware and software. For motoneurons caudal to the transection site there was a significant decrease in the mean absorbance value for NADH tetrazolium reductases which declines from 0.28 a.u. (arbitrary units) in control animals to 0.23 a.u. in cordotomized animals. However, no significant changes were detected in the activities of glucose-6-phosphate and succinate dehydrogenases. The cross-sectional area of the motoneuronal cell body was not affected by cordotomy. The decrease by around 20% in overall neuronal activity, as expressed by NADH tetrazolium reductase activity, might be expected from the decline in body motility that follows cordotomy. Changes in SDH and G6PDH activities would also be expected to follow this surgery, but none were seen, perhaps because they are compensated for by changes in neuronal metabolism that result from deafferentation.

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Detection of metabolic changes in hepatocytes by quantitative cytochemistry

et al. 1986

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Enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier

Cited by 17 publications

References 20 publications

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

Changes in enzyme histochemical profiles of identified spinal motoneurons of the European eel,Anguilla anguilla, following cordotomy

Detection of metabolic changes in hepatocytes by quantitative cytochemistry

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