Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

C. E. Blanco, Gary C Sieck, V. R. Edgerton

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

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 monoformazin which occurred with PMS and cyanide. This procedure was shown to meet various criteria established for the quantification of histochemical reactions.

Original languageEnglish (US)
Pages (from-to)230-243
Number of pages14
JournalHistochemical Journal
Volume20
Issue number4
StatePublished - 1988
Externally publishedYes

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Succinate Dehydrogenase
Azides
Skeletal Muscle Fibers
Methylphenazonium Methosulfate
Cyanides
Oxidoreductases
Formazans
Nitroblue Tetrazolium
Electron Transport Complex IV
Electrons
Muscles

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres. / Blanco, C. E.; Sieck, Gary C; Edgerton, V. R.

In: Histochemical Journal, Vol. 20, No. 4, 1988, p. 230-243.

Research output: Contribution to journalArticle

Blanco, C. E. ; Sieck, Gary C ; Edgerton, V. R. / Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres. In: Histochemical Journal. 1988 ; Vol. 20, No. 4. pp. 230-243.
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