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PMID: 3209423 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres.

The Histochemical journal ·Vol. 20 ·No. 4 ·1988-04-00 ·Pages 230-43

Blanco CE, Sieck GC, Edgerton VR

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

MeSH Terms
Animals Azides Azo Compounds/analysis Cats Cyanides Histocytochemistry Image Processing, Computer-Assisted Methylphenazonium Methosulfate/analogs & derivatives Muscles/enzymology,ultrastructure Nitroblue Tetrazolium/analysis Succinate Dehydrogenase/metabolism
Chemicals
Azides Azo Compounds Cyanides diformazan dye 1-methoxyphenazine Nitroblue Tetrazolium Methylphenazonium Methosulfate Succinate Dehydrogenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Blanco C E
Department of Anatomy, University of California, Los Angeles 90024.
Sieck G C
Edgerton V R
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32 references, click to expand
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Article Info
Journal
The Histochemical journal
Abbr.
Histochem J
ISSN
0018-2214
DOI
10.1007/BF01747468
Published
1988-04-00
Pages
230-43
Language
English
Region
Netherlands
NLM ID
0163161
Subset
IM
Grants
NHLBI NIH HHS · HL34817 · United States
NHLBI NIH HHS · HL37680 · United States
NINDS NIH HHS · NS1633 · United States
External Links
PubMed source DOI: 10.1007/BF01747468
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