Abstract
In the present study a histochemical method demonstrating the activity of dehydrogenase systems was developed for electron microscopy, utilizing potassium tellurite as the hydrogen or electron acceptor. This reagent was used intravitally (intravenously, intraperitoneally, or intraluminally in hollow organs) or supravitally on small blocks of tissue for the demonstration of endogenous dehydrogenase activity. Blocks of tissue which had been frozen and thawed or which had been washed in 0.44 M sucrose to prevent endogenous activity, were used to demonstrate the activity of the succinic dehydrogenase system. In the latter case, the incubating medium contained tellurite, succinate, phosphate buffer, sucrose, and activators. The incubation was as performed either aerobically (with or without the addition of potassium cyanide) or anaerobically. The specificity and the enzymatic nature of the reactions were ascertained by appropriate control experiments. Reduced tellurite, the end product of this histochemical reaction, could be visualized in thin sections of osmium tetroxide-fixed, methacrylate-embedded tissues as crystals or fine particulate deposits of high density, localized on, or in close relationship to mitochondrial membranes. The results of these experiments are demonstrated, utilizing heart muscle (rat) as the source of the enzyme systems.
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- CLELAND K. W., SLATER E. C. Respiratory granules of heart muscle. Biochem J. 1953 Mar;53(4):547–556. doi: 10.1042/bj0530547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GODDARD J. W., SELIGMAN A. M. Intracellular topography of succinic dehydrogenase in the thyroid of the albino rat. Anat Rec. 1952 Mar;112(3):543–559. doi: 10.1002/ar.1091120307. [DOI] [PubMed] [Google Scholar]
- HOFFMAN-OSTENHOF O. Suggestions for a more rational classification and nomenclature of enzymes. Adv Enzymol Relat Subj Biochem. 1953;14:219–220. doi: 10.1002/9780470122594.ch7. [DOI] [PubMed] [Google Scholar]
- PADYKULA H. A. The localization of succinic dehydrogenase in tissue sections of the rat. Am J Anat. 1952 Jul;91(1):107–145. doi: 10.1002/aja.1000910104. [DOI] [PubMed] [Google Scholar]
- PALADE G. E. An electron microscope study of the mitochondrial structure. J Histochem Cytochem. 1953 Jul;1(4):188–211. doi: 10.1177/1.4.188. [DOI] [PubMed] [Google Scholar]
- PALADE G. E. The fine structure of mitochondria. Anat Rec. 1952 Nov;114(3):427–451. doi: 10.1002/ar.1091140304. [DOI] [PubMed] [Google Scholar]
- PORTER K. R., PALADE G. E. Studies on the endoplasmic reticulum. III. Its form and distribution in striated muscle cells. J Biophys Biochem Cytol. 1957 Mar 25;3(2):269–300. doi: 10.1083/jcb.3.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROSA C. G., VELARDO J. T. Histochemical demonstration of succinic dehydrogenase activity in tissue sections by a modified technique. J Histochem Cytochem. 1954 Mar;2(2):110–114. doi: 10.1177/2.2.110. [DOI] [PubMed] [Google Scholar]
- RUTENBURG A. M., WOLMAN M., SELIGMAN A. M. Comparative distribution of succinic dehydrogenase in six mammals and modification in the histochemical technic. J Histochem Cytochem. 1953 Jan;1(1):66–81. doi: 10.1177/1.1.66. [DOI] [PubMed] [Google Scholar]
- SELIGMAN A. M., RUTENBURG A. M. The histochemical demonstration of succinic dehydrogenase. Science. 1951 Mar 23;113(2934):317–320. doi: 10.1126/science.113.2934.317-a. [DOI] [PubMed] [Google Scholar]
- SIEKEVITZ P., WATSON M. L. Cytochemical studies of mitochondria. II. Enzymes associated with a mitochondrial membrane fraction. J Biophys Biochem Cytol. 1956 Nov 25;2(6):653–669. doi: 10.1083/jcb.2.6.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WACHSTEIN M. Reduction of potassium tellurite by living tissues. Proc Soc Exp Biol Med. 1949 Oct;72(1):175–178. doi: 10.3181/00379727-72-17369. [DOI] [PubMed] [Google Scholar]
- WATSON M. L. The nuclear envelope; its structure and relation to cytoplasmic membranes. J Biophys Biochem Cytol. 1955 May 25;1(3):257–270. doi: 10.1083/jcb.1.3.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WITTER R. F., WATSON M. L., COTTONE M. A. Morphology and ATP-ase of isolated mitochondria. J Biophys Biochem Cytol. 1955 Mar;1(2):127–138. doi: 10.1083/jcb.1.2.127. [DOI] [PMC free article] [PubMed] [Google Scholar]