The use of ferricyanide for the light and electron microscopic demonstration of succinic dehydrogenase activity

Kerpel‐Fronius, S.; Hajós, F.

doi:10.1007/bf00304258

Cited by 97 publications

(13 citation statements)

References 23 publications

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“…The method of KERPEL-FRONIUS and HAJOS (1968) was employed for cytochemical detection of the succinic dehydrogenase activity in the slug. The incubating medium WAS prepared by mixing the solutions in the following order: 3 m10.5 M sodium-potassium tartrate dissolved in 0.1 M Sorensen's phosphate buffer (pH 7.4); 0.35 ml 0.3 M CuS04; 0.8 ml 0.1 M Sorensen's phosphate buffer (pH 7.4); 0.65 ml 1 M sodium succinate; 0.2 ml 0.05 M potassium ferricyanide.…”

Section: Cytochemical Methodsmentioning

confidence: 99%

Formation of a Prespore Specific Structure From a Mitochondrion During Development of the Cellular Slime Mold Dictyostelium Discoideum

Maeda

1971

Dev Growth Differ

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The origin of a unique vacuole (PSV), which was specifically present in the prespore cell of the cellular slime mold Dictyostelium discoideum, was investigated electronmicroscopically. A considerable number of PSV-mitochondrion complexes was found in the intermediate fraction between a pure PSV and a pure mitochondria fractions, which were obtained by isopicnic centrifugation of cellular components of the prespore cell. Similar complexes were also observed in the differentiating prespore cells. Furthermore, the activity of succinic dehydrogenase, a typical mitochondria1 enzyme was found cytochemically to be localized in the PSV as well as in mitochondria. From these results, it was concluded that the PSV was formed from the mitochondrion through some intermediate steps.A vegetative amoeba of Dictyostelium discoideum grows and multipIies independently, feeding on bacteria. Some time after ceasing to feed, amoebae stream together to collecting points (aggregation stage). The cell aggregate thus formed initially crects itself (papilla stage), but later lies down on the substratum in the shape of a slug. The slug migrates for a period of time (migration stage), until it rises into the air to form a fruiting body composed of a mass of spores and a supporting cellular stalk (culmination stage).During culmination, the anterior (prestalk) cells of the slug differentiate into stalk cells, while the posterior (prespore) cells into spores. An electronmicroscopic study has revealed that a unique vacuole named PSV is specifically present in the cytoplasm of the prespore cells, but not in the prestalk cells (MAEDA and TAKHJCHI, 1969: HOHL andHAMAMOTO, 1969). It has also been demonstrated that the vacuole (PSV) is excreted from the prespore cell during its cytodifferentiation into the spore, and that the contents constitute the spore coat (HOHL and HAMAMOTO, 1969: MAEDA, 197 1 ).The PSV was a sole structure thus far found that showed discrete localization This paper is dedicated to Prof. Emer. Tadao Sato of Nagoya University for his seventieth birthday.

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Section: Cytochemical Methodsmentioning

confidence: 99%

Formation of a Prespore Specific Structure From a Mitochondrion During Development of the Cellular Slime Mold Dictyostelium Discoideum

Maeda

1971

Dev Growth Differ

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“…As documented by several authors including ourselves (13)(14)(15)(16)(17)(18)(19), the precipitates are due to the very rapid acceptance of electrons by ferricyanide which is reduced to ferrocyanide. In turn, this molecule is captured by copper at the site of enzyme activity.…”

Section: Resultsmentioning

confidence: 90%

Aging-like alterations of SDH activity in Purkinje cell mitochondria of adult vitamin-E deficient rats

Fattoretti

Bertoni–Freddari

Casoli

et al. 2001

AGE

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The ultrastructural features of perikaryal mitochondria positive to the copper ferrocyanide cytochemical reaction due to SDH activity were investigated in Purkinje cells of adult rats fed a vitamin E (a-tocopherol) deficient diet (AVED) for 11 months. The mitochondrial volume fraction (volume density: Vv), the number of organelles/iJm 3 of tissue (numeric density: Nv) and their average volume (V) were estimated by computer-assisted morphometry. The data obtained were compared with our previous results on 3, 12 and 24 month-old normally fed rats. In a comparison with age-matched controls, AVED animals showed significant decreases of the three morphometric parameters taken into account. These reductions were also observed in old, normally fed rats vs. the young and adult groups, but in AVED rats Vv and V decreased to a higher extent. In adult control animals, the percent of larger organelles (0.32 pm 3 >) decreases to less than 1%. Vitamin E deficiency resulted in a steeper reduction of this fraction of organelles, i.e. only 0.5% in the 0.24 -0.32 IJm 3 size range accounted for the largest mitochondria in the AVED group. Taken together, these data document a significant impairment of mitochondrial efficiency in old and AVED rats. We interpret these findings to support that the underlying processes of aging and vitamin E deficiency may share common mechanisms. Considering the antioxidant action of a-tocopherol and the SDH role in cellular bioenergetics, inadequate protection from free radical attacks appears to represent an important determinant in the age-related decline of the mitochondrial metabolic competence.

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“…The results of the present examination of the ultracytochemical localization of LDH demonstrated by means of the copper ferrocyanide method (15,23,34) indicate that electron-opaque reaction product, copper ferrocyanide, (the latter denoting the site of the enzymatic activity), was found in the muscle cell membrane, the pinocytotic vesicle, the ground substance of the sarcoplasm, the T-tubules of vesicular triads and in the mitochondrial cristae, and also on the inner and outer membrane of mitochondria. In addition, LDH reaction products obtained with the A and/or B incubation medium, containing sodium potassium tartrate and sodium citrate as the chelator respectively, were detected in the same localization.…”

Section: Resultsmentioning

confidence: 91%

Ultracytochemical Study of Lactate Dehydrogenase Activity in the Tail Muscles of Anuran Tadpoles During Metamorphosis

Sasaki

1979

Acta Histochem. Cytochem

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Tail muscles of Rana catesbeiana and Xenopus laevis during metamorphosis were investigated ultracytochemically for lactate dehydrogenase (LDH). By means of the copper ferrocyanide method, LDH reaction products were found in the mitochondria, the muscle cell membrane, pinocytotic vesicle, T-tubules of vesicular triads and the sarcoplasm of the two types of muscle fibers (red and white fibers).The localization of the LDH reaction products which are described and discussed, suggests a correlation with the histochemical and biochemicalproperties of the anuran tadpole tail during metamorphosis.Light microscope histochemical studies of the localization of lactate dehydrogenase (LDH) activity have suggested that the enzyme in the two different types of striated muscle fibers of the tail of the anuran tadpole, viz. the small red peripheral muscle fiber (p.m. fiber) and the inner white muscle fiber (i.m. fiber), is localized in the sarcoplasm and the sarcoplasmic reticulum (25). Fahimi and Karnovsky (6) reported the ultrastructural localization of the LDH reaction products in the white skeletal muscle of rabbit to be in the sarcoplasmic reticulum. In the cardiac muscle of the rat, Wenzel and Behrisch (34) have found that LDH reaction products were localized in the mitochondria. Baba et al. (2) reported that the reaction products were found in the sarcoplasmic reticulum, in the ground substance of the sarcoplasm and in the mitochondria of rat muscles.A previous investigation of the inner white muscle fibers of the tadpole tail reported LDH ultrastructural localization in Rana catesbeiana, suggesting an LDH deposition of some electron-opaque materials in association with the elements of the sarcoplasmic reticulum (25).Since the red p.m. fibers and white i.m. fibers differ in their pattern of LDH isozymes and subcellular architecture (26), it is possible to conceive that isozymes of LDH have a specific ultracytochemical localization. The present study reports on the localization areas of LDH reaction products and the change in the p.m. and i.m. fibers of the anuran tadpole tail during degeneration. MATERIALS AND METHODSThe tadpoles of Rana catesbeiana used in this study were obtained from their 7

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The use of ferricyanide for the light and electron microscopic demonstration of succinic dehydrogenase activity

Cited by 97 publications

References 23 publications

Formation of a Prespore Specific Structure From a Mitochondrion During Development of the Cellular Slime Mold Dictyostelium Discoideum

Formation of a Prespore Specific Structure From a Mitochondrion During Development of the Cellular Slime Mold Dictyostelium Discoideum

Aging-like alterations of SDH activity in Purkinje cell mitochondria of adult vitamin-E deficient rats

Ultracytochemical Study of Lactate Dehydrogenase Activity in the Tail Muscles of Anuran Tadpoles During Metamorphosis

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