Changes in the cotyledons of Cucurbita maxima during germination. II. Development of mitochondrial function

1970 ◽  
Vol 48 (12) ◽  
pp. 2233-2240 ◽  
Author(s):  
John N. A. Lott ◽  
Paul Castelfranco
Keyword(s):  
Cytochrome Oxidase ◽  
Mitochondrial Function ◽  
Respiratory Activity ◽  
Succinic Dehydrogenase ◽  
Embryo Axis ◽  
Cucurbita Maxima ◽  
Tissue Slices ◽  
Electron Microscopic ◽  
Fresh Tissue ◽  
Microscopic Studies

Mitochondrial respiration in squash cotyledons was followed during germination. Cytochrome oxidase and succinoxidase activities were determined in isolated mitochondrial preparations; tissue localizations of cytochrome oxidase and succinic dehydrogenase were determined histochemically in fresh tissue slices. The respiratory activities differed in light- and dark-germinated plants. In light-grown plants, the respiratory activity reached a peak at 3 days, when the root and the stem were actively growing, and then declined markedly as the tissue became photosynthetic. During the peak, the respiratory activity was present in all the cotyledon tissues; in the foliaceous cotyledons the respiratory activity was localized in the veins. In dark-grown plants the peak of respiratory activity extended over several days and the respiratory rate remained high throughout the cotyledon's life. This broad peak of activity may be related to the mobilization of storage materials for the growth of the embryo axis. In dark-germinated plants, the respiratory activity was widespread throughout the cotyledonary tissues during the peak of activity; the activity became vein localized by 8 days. Electron microscopic studies of cotyledon tissues showed the presence of many mitochondrial profiles in the veinlet regions.

scholarly journals ULTRASTRUCTURAL DEMONSTRATION OF CYTOCHROME OXIDASE ACTIVITY BY THE NADI REACTION WITH OSMIOPHILIC REAGENTS

1967 ◽  
Vol 34 (3) ◽  
pp. 787-800 ◽  
Author(s):  
Arnold M. Seligman ◽  
Robert E. Plapinger ◽  
Hannah L. Wasserkrug ◽  
Chandicharan Deb ◽  
Jacob S. Hanker
Keyword(s):  
Cytochrome Oxidase ◽  
Oxidase Activity ◽  
Tetrazolium Salt ◽  
Tissue Slices ◽  
Fresh Tissue ◽  
Fixed Tissue ◽  
Cytochrome Oxidase Activity ◽  
Renal Tubular Cells ◽  
Renal Tubular ◽  
Rat Tissue

A new method for the subcellular and cytochemical demonstration of cytochrome oxidase has been developed with the introduction of N-benzyl-p-phenylenediamine (BPDA) and the discovery that indoanilines are osmiophilic. These indoanilines produced upon oxidation of BPDA in the presence of naphthols are highly colored compounds that yield electron-opaque coordination polymers of osmium (osmium black) that are amorphous, insoluble in water, and in organic solvents. The best methods for preparing rat tissue were in decreasing order: fixation in formaldehyde solution, fresh tissue slices, and frozen sections of fresh or fixed tissue. Ultrathin sections were counterstained by bridging with the thiocarbohydrazide-osmium tetroxide (T-O) procedure for enhancing underlying membranous structures. Cytochrome oxidase activity was noted primarily in mitochondria and occasionally in sarcotubules of heart, in mitochondria and occasionally in infoldings of the plasma membrane of renal tubular cells, and in mitochondria and, to a great extent, in endoplasmic reticulum of hepatic cells. Cytochrome oxidase activity produced deposits in droplet form, whereas dehydrogenase activity resulted in uniform staining of mitochondrial cristae, as recently demonstrated with an osmiophilic tetrazolium salt. Even more recently we have succeeded in demonstrating cytochrome oxidase activity in nondroplet staining on mitochondrial cristae with an osmiophilic benzidine-type reagent that apparently polymerizes upon oxidation (to be published later).

scholarly journals NONDROPLET ULTRASTRUCTURAL DEMONSTRATION OF CYTOCHROME OXIDASE ACTIVITY WITH A POLYMERIZING OSMIOPHILIC REAGENT, DIAMINOBENZIDINE (DAB)

1968 ◽  
Vol 38 (1) ◽  
pp. 1-14 ◽  
Author(s):  
Arnold M. Seligman ◽  
Morris J. Karnovsky ◽  
Hannah L. Wasserkrug ◽  
Jacob S. Hanker
Keyword(s):  
Cytochrome Oxidase ◽  
Respiratory Chain ◽  
Oxidase Activity ◽  
Oxidative Polymerization ◽  
Succinic Dehydrogenase ◽  
Inner Mitochondrial Membrane ◽  
Electron Microscopic ◽  
Cytochrome Oxidase Activity ◽  
Outer Compartment ◽  
Electron Microscopes

A new method for demonstrating cytochrome oxidase activity, based upon the oxidative polymerization of 3,3'-diaminobenzidine (DAB) to an osmiophilic reaction product, has improved the localization of this enzyme over methods based upon the Nadi reaction, in both the light and electron microscopes. The reaction product occurs in nondroplet form, which more accurately delineates the localization of cytochrome oxidase in mitochondria of heart, liver, and kidney. In electron microscopic preparations the excess reaction product is found to overflow into the intracristate spaces and into the outer compartment between inner and outer limiting mitochondrial membranes. This finding suggests that the enzymatic activity of cytochrome c is located on the inner surface of the intracristate space which is the outer surface of the inner mitochondrial membrane. Succinic dehydrogenase activity has also been located at this site by using an osmiophilic ditetrazolium salt, TC-NBT. Considered together, the sites of reactivity of both parts of the respiratory chain have implications for the chemiosomotic hypothesis of Mitchell who suggests a mechanism of energy conservation during electron transport in the respiratory chain of the mitochondrion.

Intracellular location of enzymes in Rhodopseudomonas spheroides

1968 ◽  
Vol 170 (1020) ◽  
pp. 319-329 ◽  
Keyword(s):  
Cell Wall ◽  
Cytoplasmic Membrane ◽  
Succinic Dehydrogenase ◽  
Particulate Material ◽  
Zinc Protoporphyrin ◽  
Differential Centrifugation ◽  
Electron Microscopic ◽  
Intracellular Location ◽  
Microscopic Studies ◽  
Rhodopseudomonas Spheroides

By differential centrifugation of extracts of pigmented Rhodopseudomonas spheroides a number of constituents, phospholipid and lipid ornithine, and enzymes, zinc protoporphyrin chelatase, succinic dehydrogenase and S-adenosylmethionine-magnesium protoporphyrin methyltransferase, have been found to be associated both with chromatophores and with non-pigmented particulate material. These components are present in both types of material at about the same level. In extracts of non-pigmented organisms the particulate material contains some of the above components, but others are only present in low amounts. The subcellular structures present in the particulate material—ribosomes, cell wall and cytoplasmic membrane—have only been partially separated but, by comparing the distribution of the components listed above with those of known components of ribosomes and cell wall, it is probable that they are associated with cytoplasmic membrane. These studies suggest that the cytoplasmic membrane, apart from lacking the photosynthetic pigments, has a composition similar to that of chromatophores. The data are consistent with the conclusion drawn from electron microscopic studies that chromatophores are derived by invagination of the cytoplasmic membrane.

scholarly journals THE METABOLISM OF THE KIDNEY IN EXPERIMENTAL RENAL HYPERTENSION

1945 ◽  
Vol 82 (4) ◽  
pp. 227-240 ◽  
Author(s):  
Sigwin B. Raska
Keyword(s):  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Amine Oxidase ◽  
Renal Hypertension ◽  
Succinic Dehydrogenase ◽  
Inhibitory Effect ◽  
Acid Oxidase ◽  
Tissue Slices ◽  
Amino Acid Oxidase ◽  
Experimental Renal Hypertension

These investigations are part of an attempt to study and interpret the intermediary metabolism of the kidneys in experimental renal hypertension. Hypertension was produced in dogs by the clamping procedure of Goldblatt and associates or by the silk perinephritis method of Page. Enzymatic studies were made by means of Warburg's manometric method. Cytochrome c was in addition determined spectrophotometrically. Tissue slices, homogenized tissue, and tissue extracts were used. A study of the cytochrome c concentration and the activities of the cytochrome oxidase and succinic dehydrogenase systems of kidneys from normal dogs and dogs with experimental renal hypertension was made. It was found that the cytochrome c concentration and the activities of the cytochrome oxidase and succinic dehydrogenase systems were markedly lower in the kidney slices and in the tissue suspensions from hypertensive dogs. Tissue suspensions and extracts of kidneys from hypertensive dogs showed an inhibitory effect on the activity of the cytochrome oxidase and succinic dehydrogenase, and the amine oxidase systems. Renin preparations also showed a marked inhibitory effect on the activities of cytochrome oxidase, succinic dehydrogenase, l-amino acid oxidase, and amine oxidase systems. A significant increase was found in the kidney of dogs whose other kidney had been removed or subjected to Goldblatt's or Page's technique in the activities of the cytochrome-cytochrome oxidase system, the succinic dehydrogenase system, and in the concentration of nucleotide-bound phosphorus, of flavin-adenine dinucleotide, and of the nicotinamide-adenine dinucleotides (coenzymes I and II). From the results of these studies it can be concluded that an increase in the concentration and activity of the respiratory enzymes precedes hypertrophy of the kidney. This can be explained by the assumption that an increase in the activity of the respiratory biocatalysts acts as a stimulus for cell growth and multiplication.

Electron microscopic studies on ultrathin frozen sections of lactating mouse mammary gland

1978 ◽  
Vol 36 (2) ◽  
pp. 46-47
Author(s):  
Jan Zarzycki ◽  
Joseph Szroeder
Keyword(s):  
Mammary Gland ◽  
Protein Denaturation ◽  
Chemical Constituents ◽  
Freeze Substitution ◽  
Electron Microscopic Examination ◽  
Mouse Mammary Gland ◽  
Electron Microscopic ◽  
Preparation Methods ◽  
Microscopic Studies ◽  
Ultrathin Frozen Sections

The mammary gland ultrastructure in various functional states is the object of our investigations. The material prepared for electron microscopic examination by the conventional chemical methods has several limitations, the most important are the protein denaturation processes and the loss of large amounts of chemical constituents from the cells. In relevance to this,one can't be sure about a degree the observed images are adequate to the realy ultrastructure of a living cell. To avoid the disadvantages of the chemical preparation methods,some autors worked out alternative physical methods based on tissue freezing / freeze-drying, freeze-substitution, freeze-eatching techniqs/; actually the technique of cryoultraraicrotomy,i,e.cutting ultrathin sections from deep frozen specimens is assented as a complete alternative method. According to the limitations of the routine plastic embbeding methods we were interested to analize the mammary gland ultrastructure during lactation by the cryoultramicrotomy method.

Early Development of Drug-Induced Hepatic Necrosis

1971 ◽  
Vol 29 ◽  
pp. 576-577
Author(s):  
F. G. Zaki ◽  
E. Detzi ◽  
C. H. Keysser
Keyword(s):  
Early Development ◽  
Hepatic Necrosis ◽  
Screening Tests ◽  
Dense Matrix ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
Drug Induced ◽  
Hepatocellular Damage ◽  
Sprague Dawley Rats ◽  
Microscopic Studies

This study represents the first in a series of investigations carried out to elucidate the mechanism(s) of early hepatocellular damage induced by drugs and other related compounds. During screening tests of CNS-active compounds in rats, it has been found that daily oral administration of one of these compounds at a dose level of 40 mg. per kg. of body weight induced diffuse massive hepatic necrosis within 7 weeks in Charles River Sprague Dawley rats of both sexes. Partial hepatectomy enhanced the development of this peculiar type of necrosis (3 weeks instead of 7) while treatment with phenobarbital prior to the administration of the drug delayed the appearance of necrosis but did not reduce its severity.Electron microscopic studies revealed that early development of this liver injury (2 days after the administration of the drug) appeared in the form of small dark osmiophilic vesicles located around the bile canaliculi of all hepatocytes (Fig. 1). These structures differed from the regular microbodies or the pericanalicular multivesicular bodies. They first appeared regularly rounded with electron dense matrix bound with a single membrane. After one week on the drug, these vesicles appeared vacuolated and resembled autophagosomes which soon developed whorls of concentric lamellae or cisterns characteristic of lysosomes (Fig. 2). These lysosomes were found, later on, scattered all over the hepatocytes.

Electron Microscopic identification of Pre-B Lymphocytes in the Bone Marrow of a Patient with Chronic Myelocytic Leukemic in Blast Crisis

1982 ◽  
Vol 40 ◽  
pp. 346-347
Author(s):  
T. Mullin ◽  
G. Yee ◽  
M. Aheam ◽  
J. Trujillo
Keyword(s):  
Blast Crisis ◽  
Immunoglobulin M ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Transformation Theory ◽  
Electron Microscopic ◽  
Chemotherapeutic Sensitivity ◽  
Microscopic Studies ◽  
Hematopoietic Precursor ◽  
Lymphoblastic Transformation ◽  
B Lineage

There have been numerous reports in the current literature suggesting that hematopoietic precursor cells in some human chronic myelocytic leukemias (CML) undergo lymphoblastic transformation at the time of the acute blast crisis (BC) stage. The primary evidence offered in support of this transformation theory--lymphoblastic appearing morphology, increased terminal deoxynucleotidyl transferase (TdT) activity, and chemotherapeutic sensitivity to vincristine and prednisone--has been indirect, however, since these features may occur in nonlymphoid cells. More direct support for the Pre-B lineage of these cells has recently been provided by immunofluorescent light microscopic studies demonstrating the presence of intracytoplasmic immunoglobulin M (IgM) in these CML-BC cells.

Origin of Hepatic Nuclear Inclusion Bodies

1973 ◽  
Vol 31 ◽  
pp. 426-427
Author(s):  
F. G. Zaki ◽  
J. A. Greenlee ◽  
C. H. Keysser
Keyword(s):  
Inclusion Bodies ◽  
Storage Disease ◽  
Glycogen Storage ◽  
Nutritional Deficiencies ◽  
Nuclear Inclusion ◽  
Electron Microscopic ◽  
Human Liver Cells ◽  
Microscopic Studies ◽  
Per Se ◽  
Experimental Liver

Nuclear inclusion bodies seen in human liver cells may appear in light microscopy as deposits of fat or glycogen resulting from various diseases such as diabetes, hepatitis, cholestasis or glycogen storage disease. These deposits have been also encountered in experimental liver injury and in our animals subjected to nutritional deficiencies, drug intoxication and hepatocarcinogens. Sometimes these deposits fail to demonstrate the presence of fat or glycogen and show PAS negative reaction. Such deposits are considered as viral products.Electron microscopic studies of these nuclei revealed that such inclusion bodies were not products of the nucleus per se but were mere segments of endoplasmic reticulum trapped inside invaginating nuclei (Fig. 1-3).

Novel methods for demonstrating osseointegration of hydroxylapatite-coated titanium hip prostheses

1991 ◽  
Vol 49 ◽  
pp. 34-35 ◽  
Author(s):  
P. Frayssinet ◽  
J. Hanker ◽  
D. Hardy ◽  
B. Giammara
Keyword(s):  
Hip Prosthesis ◽  
Ethanol Concentration ◽  
Bone Matrix ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Hip Prostheses ◽  
Cellular Inclusions ◽  
Microscopic Studies ◽  
Diamond Saw ◽  
Plasma Sprayed

Prostheses implanted in hard tissues cannot be processed for electron microscopic examination or microanalysis in the same way as those in other tissues. For these reasons, we have developed methods allowing light and electron microscopic studies as well as microanalysis of the interface between bone and a metal biomaterial coated by plasma-sprayed hydroxylapatite(HA) ceramic.An HA-coated titanium hip prosthesis (Corail, Landos, France), which had been implanted for two years, was removed after death (unrelated to the orthopaedic problem). After fixation it was dehydrated in solutions of increasing ethanol concentration prior to embedment in polymethylmethacrylate(PMMA). Transverse femur sections were obtained with a diamond saw and the sections then carefully ground to a thickness of 200 microns. Plastic-embedded sections were stained for calcium with a silver methenamine modification of the von Kossa method for calcium staining and coated by carbon. They have been examined by back-scatter SEM on an ISI-SS60 operated at 25 KV. EDAX has been done on cellular inclusions and extracellular bone matrix.

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