Isolation of intramitochondrial helical filaments appearing in outer compartment of mitochondria

1995 ◽  
Vol 241 (2) ◽  
pp. 149-154 ◽  
Author(s):  
Hiroyuki Sasaki ◽  
Susumu Kurioka ◽  
Hiroyuki Fukata ◽  
Takao Ohoki ◽  
Hisako Arai ◽  
...  
Keyword(s):  
Outer Compartment

scholarly journals The ultrastructural cytochemistry of lactic dehydrogenase, succinic dehydrogenase, dihydro-nicotinamide adenine dinucleotide diaphorase and cytochrome oxidase activities in hair cell mitochondria of the guinea pig cochlea.

1975 ◽  
Vol 23 (3) ◽  
pp. 216-234 ◽  
Author(s):  
G J Spector
Keyword(s):  
Hair Cell ◽  
Cytochrome Oxidase ◽  
Nicotinamide Adenine Dinucleotide ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Nicotinamide Adenine ◽  
Fixation Method ◽  
Inner Mitochondrial Membrane ◽  
Tetrazolium Chloride ◽  
Outer Compartment

The use of cinnamyl nitroblue tetrazolium chloride (DS-NBT) in dehydrogenase experiments (lactic dehydrogenase, succinic dehydrogenase, nicotinamide adenine dinucleotide diaphorase) and 3,3'-diaminobenzidine tetrahydrochloride (DAB) in cytochrome oxidase experiments indicated that mitochondrial oxidoreduction reactions from nicotinamide adenine dinucleotide to cytochrome oxidase are located on the inner mitochondrial membrane in the outer compartment and the intracristate spaces. These reactions behave according to the chemiosmotic hypothesis. The cochlear hair cell mitochondria are cytochemically indistinguishable from free liver mitochondria. The heterogeneous mitochondrial staining pattern is related to the osmolarity of the incubation media, solubility of the enzymes and pH of the medium, but not to the fixation method.

scholarly journals A study of rapid mitochondrial structural changes in vitro by spray-freeze-etching

1978 ◽  
Vol 77 (1) ◽  
pp. 134-147 ◽  
Author(s):  
RD Lang ◽  
JR Bronk
Keyword(s):  
Time Course ◽  
Structural Changes ◽  
Rat Liver Mitochondria ◽  
Sampling Device ◽  
Rapid Sampling ◽  
Outer Compartment ◽  
Matrix Volume ◽  
The Matrix ◽  
Freeze Etching

The spray-freeze-etching technique has been used to study energy-linked mitochondrial structural changes in rat liver mitochondria incubated in vitro. The technique involves spraying the suspension of mitochondria into liquid propane at -190 degrees C, and does not require the use of cryoprotectants or chemical fixatives. The results confirmed that freshly isolated mitochondria have a condensed matrix and that this expands at the expense of the outer compartment to give the orthodox configuration when the mitochondria are incubated in a K+ medium in the presence of substrate and phosphate. Addition of adenosine diphosphate (ADP) caused a rapid shrinkage of the matrix compartment, and the time-course and extent of this shrinkage has been measured quantitatively by coupling a rapid sampling device to the spray-freezing apparatus. These data show that for orthodox mitochondria the onset of phosphorylation is accompanied by a reduction of 30% in the matrix volume in 20's, and there is no evidence that the decrease in matrical volume affects the phosphorylation efficiency. These results suggest that natural ionophores in the mitochondrial inner membrane make it permeable enough to permit a rapid readjustment of matrix volume after the addition of ADP, and that the associated ion movement does not cause uncoupling of oxidative phosphorylation.

scholarly journals ULTRASTRUCTURAL VARIATIONS IN N,N'-BIS(4-AMINOPHENYL)-N,N'-DIMETHYLETHYLENEDIAMINE (BED) OXIDASE LOCALIZATION IN RAT LIVER AND PAROTID GLAND WITH VARIATION IN LENGTH OF FIXATION

1972 ◽  
Vol 20 (12) ◽  
pp. 1024-1030 ◽  
Author(s):  
W. ALLEN SHANNON ◽  
ARNOLD M. SELIGMAN
Keyword(s):  
Endoplasmic Reticulum ◽  
Parotid Gland ◽  
Rat Liver ◽  
Rough Endoplasmic Reticulum ◽  
Secretory Granules ◽  
Terminal Oxidase ◽  
Outer Compartment ◽  
Fixation Reaction

The localization and reactivity of a terminal oxidase which oxidizes N,N'-bis(4-amino-phenyl)-N,N'-dimethylethylenediamine (BED) were studied in rat liver and parotid gland after varying the concentration of formaldehyde fixative and the length of fixation. Reaction product was observed in mitochondrial outer compartments, smooth elements of rough endoplasmic reticulum, some Golgi lamellae, perinuclear membranes and cytoplasmic membranous structures often associated with mitochondria. A reaction also occurred in the limiting membrane and, to some degree, in the material comprising the secretory granules of the parotid. The reaction in the mitochondrial outer compartment was extremely formaldehyde-sensitive. Controls in which diaminobenzidine (DAB) was substituted for BED showed reaction only in mitochondrial cristae and outer compartments, whereas controls without either reagent showed no reactivity.

Development of the larval tunic in a compound ascidian: morphogenetic events in the embryos of Distaplia occidentalis

1984 ◽  
Vol 62 (12) ◽  
pp. 2392-2400 ◽  
Author(s):  
Michael J. Cavey ◽  
Richard A. Cloney
Keyword(s):  
External Surface ◽  
Body Cavity ◽  
The Body ◽  
Ground Substance ◽  
Perivitelline Space ◽  
Outer Compartment ◽  
Test Cells ◽  
Principal Elements

The larval tunic of Distaplia occidentalis is a complex investment consisting of extracellular filaments and an amorphous ground substance. The principal elements of the tunic are two thin cuticles and two subcuticular compartments. Compacted filaments and small amounts of ground substance characterize the cuticles. Dispersed filaments and large amounts of ground substance distinguish the compartments. The cells of the embryonic epidermis apparently secrete both the filaments and the ground substance. The elements of the larval tunic differentiate sequentially over a period of approximately 2 weeks. The outer cuticle appears 3–4 days after neurulation and it gradually lifts from the surface of the epidermis as the outer compartment forms. Tapered folds of the outer cuticle and extensions of the outer compartment produce the dorsal, ventral, and posterior fins of the larva. Test cells, occupying the perivitelline space around the embryo, release multigranular "ornaments" that adhere to the external surface of the outer cuticle. The inner cuticle and the inner compartment arise during the 4 days prior to hatching. The inner compartment over the truncal epidermis expands significantly to accommodate cells which emigrate from the body cavity (hemocoel).

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.

THE SPECIFIC ACTIVITY OF PLASMA CORTISOL IN SHEEP AFTER INTRAVENOUS INFUSION OF [1,2-3H2]CORTISOL, AND ITS RELATION TO THE DISTRIBUTION OF CORTISOL

1968 ◽  
Vol 40 (1) ◽  
pp. 37-47 ◽  
Author(s):  
J. Y. F. PATERSON ◽  
F. A. HARRISON
Keyword(s):  
Intravenous Infusion ◽  
Plasma Cortisol ◽  
Specific Activity ◽  
Compartment Model ◽  
Central Compartment ◽  
Cortisol Concentration ◽  
Two Compartment Model ◽  
Outer Compartment ◽  
Constant Rate ◽  
Near Term

SUMMARY Tritium-labelled cortisol was administered to sheep by intravenous infusion at constant rate for up to 4 hr. When the infusion was stopped, [3H] cortisol disappeared rapidly from plasma and its concentration could be described by a double exponential function. There was good agreement between the results from 50 experiments on 11 sheep. In pregnant ewes, there was no noticeable difference in the rate of disappearance of [3H]cortisol from plasma until about 2 weeks before lambing, when the rate became more rapid. These data were interpreted in terms of a two-compartment model of cortisol distribution. The central compartment contains about 42 μg. cortisol and may be identical with the cortisol contained in whole blood volume. The outer compartment contains about 130 μg. cortisol; less than half of this compartment may be in intercellular fluids, partly bound to protein, and the remainder in intracellular fluids. In pregnant ewes near term there is a decrease in plasma cortisol concentration which appears to result from expansion of plasma volume. The decrease in unbound cortisol concentration probably results in a decrease in the size of the outer compartment of cortisol. This may contribute to the observed increase in the rate of disappearance of [3H] cortisol from plasma, but this change may also coincide with the initiation of secretion of cortisol by the foetus, at about 1 or 2 μg./min.

scholarly journals Sox2 and FGF20 interact to regulate organ of Corti hair cell and supporting cell development in a spatially-graded manner

2018 ◽  
Author(s):  
Lu M. Yang ◽  
Kathryn S.E. Cheah ◽  
Sung-Ho Huh ◽  
David M. Ornitz
Keyword(s):  
Hearing Loss ◽  
Hair Cell ◽  
Hair Cells ◽  
Genetic Interaction ◽  
Organ Of Corti ◽  
Supporting Cell ◽  
Cell Development ◽  
Supporting Cells ◽  
Outer Compartment ◽  
Mouse Organ

AbstractThe mouse organ of Corti develops in two steps: progenitor specification and differentiation. Fibroblast Growth Factor (FGF) signaling is important in this developmental pathway, as deletion of FGF receptor 1 (Fgfr1) or its ligand, Fgf20, leads to the loss of hair cells and supporting cells from the organ of Corti. However, whether FGF20-FGFR1 signaling is required during specification or differentiation, and how it interacts with the transcription factor Sox2, also important for hair cell and supporting cell development, has been a topic of debate. Here, we show that while FGF20-FGFR1 signaling functions during progenitor differentiation, FGFR1 has an FGF20-independent, Sox2-dependent role in specification. We also show that a combination of reduction in Sox2 expression and Fgf20 deletion recapitulates the Fgfr1-deletion phenotype. Furthermore, we uncovered a strong genetic interaction between Sox2 and Fgf20, especially in regulating the development of hair cells and supporting cells towards the basal end and the outer compartment of the organ of Corti. To explain this genetic interaction and its effects on the basal end of the organ of Corti, we provide evidence that decreased Sox2 expression delays specification, which begins at the organ of Corti apex, while Fgf20-deletion results in premature onset of differentiation, which begins near the organ of Corti base. Thereby, Sox2 and Fgf20 interact to ensure that specification occurs before differentiation towards the cochlear base. These findings reveal an intricate developmental program regulating organ of Corti development along the basal-apical axis of the cochlea.Author summaryThe mammalian cochlea contains the organ of Corti, a specialized sensory epithelium populated by hair cells and supporting cells that detect sound. Hair cells are susceptible to injury by noise, toxins, and other insults. In mammals, hair cells cannot be regenerated after injury, resulting in permanent hearing loss. Understanding genetic pathways that regulate hair cell development in the mammalian organ of Corti will help in developing methods to regenerate hair cells to treat hearing loss. Many genes are essential for hair cell and supporting cell development in the mouse organ of Corti. Among these are Sox2, Fgfr1, and Fgf20. Here, we investigate the relationship between these three genes to further define their roles in development.Interestingly, we found that Sox2 and Fgf20 interact to affect hair cell and supporting cell development in a spatially-graded manner. We found that cells toward the outer compartment and the base of the organ of Corti are more strongly affected by the loss of Sox2 and Fgf20. We provide evidence that this spatially-graded effect can be partially explained by the roles of the two genes in the precise timing of two sequential stages of organ of Corti development, specification and differentation.

Purification of the helical filaments appearing in mitochondria

1990 ◽  
Vol 48 (3) ◽  
pp. 532-533
Author(s):  
Hiroyuki Sasaki ◽  
Susumu Kurioka ◽  
Takeyuki Misawa ◽  
Teruo Suzuki
Keyword(s):  
Epithelial Cells ◽  
Helical Structure ◽  
Liver Cells ◽  
Proximal Tubules ◽  
Ethanol Ingestion ◽  
Isolated Mitochondria ◽  
Molecular Arrangement ◽  
Outer Compartment ◽  
Parenchymal Cells ◽  
The Brain

Helical filaments with characteristic morphology have been found exclusively in the outer compartment of mitochondria in the epithelial cells of renal proximal tubules of patients, and in the brain astrocytes, the ameloblasts and the epithelial cells of renal medullary tubules of the normal rats, and in the liver parenchymal cells of the rats with and without ethanol ingestion. Recently, the filaments with same helical structure found in the mitochondria of the certain parasite. Thus, the intramitochondrial helical filaments appeared widely in biological fields. However, the nature, origin and physiological significance of the helical filament are at present unknown. In the present study, to clarify the molecular arrangement of the helical filament, the filaments were purified from the mitochondriaof the liver of ethanol ingested rats in which have been confirmed to appear large amount of the helical filaments in mitochondria.Male S-D strain rats (6-week old) were used in the present study. Rat were given 30% ethanol for 90 days with oral administration. With conventional TEM observations, the helical filaments were frequently found in mitochondria of liver cells of the rat with ethanol ingestion (Fig. 1). For purification of the helical filaments, the mitochondria were isolated from ethanol ingested rats liver cells by slightly modified Bustamante et al. method. The isolated mitochondria were ruptured by osmotic breakage.

Sign in / Sign up

Export Citation Format

Share Document