scholarly journals HISTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE IN HAMSTER AND RABBIT EGGS

1965 ◽  
Vol 13 (6) ◽  
pp. 470-475 ◽  
Author(s):  
K. ISHIDA ◽  
M. C. CHANG
Keyword(s):  
Gradual Increase ◽  
Succinic Dehydrogenase ◽  
Blastocyst Stage ◽  
Histochemical Demonstration ◽  
Tetrazolium Salt ◽  
Succinic Dehydrogenase Activity ◽  
Strong Activity ◽  
Moderate Reaction ◽  
Time Required ◽  
Mammalian Eggs

Succinic dehydrogenase activity, using Nitro-blue tetrazolium salt as electron acceptor, has been demonstrated histochemically in hamster and rabbit eggs before implantation. The formazans formed as blue granules are spread throughout the cytoplasm and among vitelline granules, corresponding to the distribution of mitochondria. The intensity and distribution of these granules vary according to the developmental stage of the eggs. Incubation of hamster eggs for 1 hour produced a weekly and evenly distributed reaction in most (92%) of the unfertilized eggs, while 57% of fertilized 1-cell eggs showed a moderate reaction. Although 84% of the 2-cell eggs, 95% of the 4-cell eggs, and 98% of the 8-cell eggs showed moderate reaction, 78% of blastocysts showed strong activity. In the rabbit egg the time required to develop the reaction was about 4-6 hours for unfertilized and pronuclear eggs, 3-5 hours for 2-cell eggs, 2 hours for 32-cell eggs, 1-2 hours for morulae, and only 0.5-1 hour for blastocysts. The development of succinic dehydrogenase in mammalian eggs at the first cleavage, its gradual increase during cleavage, and its tremendous increase at the blastocyst stage have been clearly demonstrated. The importance of this enzyme activity in relation to oxygen uptake, pathways of carbohydrate metabolism, and the degeneration of eggs are discussed.

scholarly journals NONOSMIOPHILIC TETRAZOLIUM SALTS THAT YIELD OSMIOPHILIC, LIPOPHOBIC FORMAZANS FOR ULTRASTRUCTURAL LOCALIZATION OF DEHYDROGENASE ACTIVITY

1972 ◽  
Vol 20 (9) ◽  
pp. 685-695 ◽  
Author(s):  
MOSHE KALINA ◽  
ROBERT E. PLAPINGER ◽  
YOSHINOBU HOSHINO ◽  
ARNOLD M. SELIGMAN
Keyword(s):  
Dehydrogenase Activity ◽  
Osmium Tetroxide ◽  
Succinic Dehydrogenase ◽  
Ultrastructural Localization ◽  
Nitroblue Tetrazolium ◽  
Tetrazolium Salt ◽  
Succinic Dehydrogenase Activity ◽  
Tetrazolium Chloride ◽  
Tetrazolium Salts ◽  
Cytochemical Demonstration

Monotetrazolium salts were designed and prepared which incorporate a phthalhydrazide moiety to make them lipophobic and a benzothiazole moiety to make them react with osmium tetroxide after they are reduced to the corresponding formazans. The tetrazolium salts themselves do not react with osmium tetroxide under the conditions used for the cytochemical demonstration of enzyme activities. Although all of the new formazans, when dissolved in dimethylformamide, were reoxidized to the tetrazolium salts by osmium tetroxide, they were not reoxidized by osmium tetroxide when they were dissolved in tetrahydrofuran or precipitated by reduction in tissue, but gave dark complexes. One of the tetrazolium salts, 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT), was readily reduced by succinic dehydrogenase activity, gave a formazan which produced a dark osmium complex relatively rapidly and gave good localization of succinic dehydrogenase activity on the membranes of mitochondria of rat myocardial cells. Although this tetrazolium salt (BSPT) was not photoreduced by fresh cells of Elodea to stain chloroplasts as seen with nitroblue tetrazolium or distyryl nitroblue tetrazolium, the 5-p-nitrophenyl derivative of BSPT was photoreduced by chloroplasts. The preparation of BSPT and its use in demonstrating succinic dehydrogenase activity ultracytochemically is given here. The preparation and ultracytochemical use of the 5-p-nitrophenyl derivative in chloroplasts will be published later.

scholarly journals UBIQUINONE AND PHOSPHOLIPIDS AS LIMITING FACTORS IN THE HISTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE ACTIVITY

1967 ◽  
Vol 15 (2) ◽  
pp. 79-82 ◽  
Author(s):  
MOSHE WOLMAN ◽  
JOSE JAIME BUBIS
Keyword(s):  
Dehydrogenase Activity ◽  
Succinic Dehydrogenase ◽  
Histochemical Demonstration ◽  
Mouse Kidney ◽  
Limiting Factors ◽  
Succinic Dehydrogenase Activity ◽  
Similar Treatment ◽  
Kidney Liver ◽  
Cryostat Sections ◽  
Long Storage

Deposition of ubiquinone and preferably of a crude phospholipid mixture and ubiquinone on cryostat sections of mouse kidney, liver and heart increased the intensity of staining for succinic dehydrogenase. A similar treatment partly restored the activity of this enzyme which was reduced by exposure to 60°C, long storage at 37°C, oxidation and treatment with lipid solvents. The restoration was present only in the case of mild noxious treatments. It is suggested that removal or structural alteration of ubiquinone and phospholipids may limit the rate of dehydrogenase activity; more drastic treatments affect the enzyme itself so that these factors cannot exert any restorative activity.

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