The Histochemical Journal

, Volume 20, Issue 4, pp 230–243 | Cite as

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres

  • C. E. Blanco
  • G. C. Sieck
  • V. R. Edgerton
Papers

Summary

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.

Keywords

Cyanide Azide Succinic Dehydrogenase Nitroblue Tetrazolium Phenazine 

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Copyright information

© Chapman and Hall Ltd 1988

Authors and Affiliations

  • C. E. Blanco
    • 1
    • 2
    • 3
  • G. C. Sieck
    • 1
    • 2
    • 3
    • 4
  • V. R. Edgerton
    • 2
    • 3
  1. 1.Department of Anatomy, School of Medicine, Center for the Health SciencesUniversity of California at Los AngelesLos AngelesUSA
  2. 2.Department of KinesiologyUniversity of California at Los AngelesLos AngelesUSA
  3. 3.Department of Brain Research InstituteUniversity of California at Los AngelesLos AngelesUSA
  4. 4.Lung InstituteCity of Hope National Medical CenterDuarteUSA

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