scholarly journals ELECTRON MICROSCOPIC OBSERVATIONS ON THE ACCUMULATION OF DIVALENT CATIONS IN INTRAMITOCHONDRIAL GRANULES

1964 ◽  
Vol 20 (1) ◽  
pp. 95-111 ◽  
Author(s):  
Lee D. Peachey

Electron microscopic evidence is presented, from mitochondria in whole cells of toad urinary bladder and from isolated rat kidney mitochondria, indicating that the divalent cations calcium, strontium, and barium are accumulated in granules localized in the mitochondrial matrix. This accumulation occurs under conditions in which divalent ions are present in the medium bathing either whole cells or isolated mitochondria. The evidence indicates that the divalent ions are deposited on, or in a pre-existing granule, possibly in exchange for other ions. It suggests a possible role of the intramitochondrial granules in the regulation of the internal ionic environment of the mitochondrion. Certain biochemical and physiological implications of this phenomenon are discussed.

1964 ◽  
Vol 23 (2) ◽  
pp. 207-215 ◽  
Author(s):  
Mario H. Burgos ◽  
Agustin Aoki ◽  
Fabio L. Sacerdote

Direct electron microscopic evidence is reported of the ultrastructure of mitochondrial membranes and compartments in mitochondria isolated in 0.5 M sucrose from the rat kidney cortex and the experimental changes they undergo with phlorizin and ATP treatment. A heterogeneous population of mitochondria is recognized under control conditions. The mitochondria appear to be of 3 main types, normal, swollen, and contracted. Under phlorizin treatment, most of the mitochondria swell in less than 15 minutes, apparently at the expense of the matrix. Treatment with ATP, on the other hand, produces, during the same time, a marked contraction of the isolated mitochondria, with many refoldings of the inner membrane and marked increase in the electron opacity of the matrix. It is concluded from these observations that mitochondrial swelling and contraction should be related mainly to the matrix content.


Author(s):  
S. E. Coleman ◽  
J. Duggan ◽  
R. L. Hackett

Complete renal ischemia causes a series of subcellular changes including chromatin clumping, dilation of endoplasmic reticulum and condensation of mitochondrial matrix followed by swelling of the mitochondria and unfolding of the cristae (1). Most mitochondria are condensed by 15 minutes and half are condensed and half swollen by 30 minutes with the majority of the mitochondria swollen by 60 minutes of ischemia.In this study we have looked at the ability of mitochondria isolated from ischemic rat kidneys to undergo structural energy transformations in the presence of substrate and ADP. Mitochondria were isolated from the inner cortex of rat kidney where initial damage from ischemia occurs. The tissue was diced in the cold, homogenized with a Dounce homogenizer, spun twice at 600 g, and the supernatant at 15,000 g for pelleting of mitochondria.


1988 ◽  
Vol 254 (2) ◽  
pp. F191-F195
Author(s):  
M. Le Hir ◽  
U. C. Dubach

Hydrolysis of 5'-AMP by 5'-nucleotidase is a possible source of adenosine in the kidney. A renal membrane-bound ecto-5'-nucleotidase has been previously described. The present study deals with the catalytic properties of a 5'-AMP phosphohydrolase partially purified from high-speed supernatants of rat kidney homogenates. It exhibits phosphatase activity toward 5'-AMP, 5'-IMP, and 5'-GMP, but not toward 2'- and 3'-AMP and corresponds therefore to a 5'-nucleotidase. The hydrolysis of 5'-AMP by the soluble 5'-nucleotidase requires divalent cations. Maximal activity is reached with 10 microM of either Mn2+ or Co2+, whereas half-maximal activity is obtained with approximately 400 microM Mg2+. The soluble 5'-nucleotidase exhibits Michaelis-Menten kinetics with a Km of 9.5 microM for 5'-AMP. In the presence of 1 mM of free Mg2+, physiological concentrations of ATP provoke an increase of the Km for 5'-AMP and a decrease of Vmax. An increase of the pH of 0.4 units in the pH range 6.4-7.4 roughly doubles the rate of hydrolysis of 5'-AMP. The effects of ATP and of the pH are compatible with a role of the renal soluble 5'-nucleotidase in the hydrolysis of 5'-AMP and in the production of adenosine during hypoxia.


1978 ◽  
Vol 56 (1) ◽  
pp. 23-28 ◽  
Author(s):  
Surinder Cheema-Dhadli ◽  
Mitchell L. Halperin

Since glutamine enters rat kidney mitochondria without exchange for an anion, the exit of its carbon skeleton must involve the dicarboxylate anion transporter (malate – inorganic phosphate) for ammoniagenesis to proceed. Therefore, this important mitochondrial anion transporter was studied in isolated renal cortex mitochondria. The phosphate concentration required for half-maximal rates of malate exit from renal cortex mitochondria of normal rats was 1.0 mM. This value was not decreased in renal cortex mitochondria from rats with chronic metabolic acidosis. The maximum velocity of the dicarboxylate transporter was not increased in renal cortex mitochondria from these acidotic rats. These kinetic parameters were similar in liver mitochondria. There was no acute activation of the dicarboxylate carrier when the incubation medium pH was lowered. Thus, there is no demonstrable activation of the dicarboxylate anion transporter in kidney cortex mitochondria of the rat with chronic metabolic acidosis. The significance of these results with respect to the regulation of renal ammoniagenesis is discussed.


1962 ◽  
Vol 12 (1) ◽  
pp. 57-78 ◽  
Author(s):  
L. E. Roth ◽  
E. W. Daniels

Dividing nuclei from the giant ameba Pelomyxa carolinensis were fixed in osmium tetroxide solutions buffered with veronal acetate to pH 8.0. If divalent cations (0.002 M calcium, magnesium, or strontium as chlorides) were added to the fixation solution, fibrils that are 14 mµ in diameter and have a dense cortex are observed in the spindle. If the divalent ions were omitted, oriented particles of smaller size are present and fibrils are not obvious. The stages of mitosis were observed and spindle components compared. Fibrils fixed in the presence of calcium ions are not so well defined in early metaphase as later, but otherwise have the same diameter in the late metaphase, anaphase, and early telophase. Fibrils are surrounded by clouds of fine material except in early telophase, when they are formed into tight bundles lying in the cytoplasm unattached to nuclei. Metaphase and anaphase fibrils fixed without calcium ions are less well defined and are not observably different from each other. The observations are consistent with the concept that spindle fibrils are composed of polymerized, oriented protein molecules that are in equilibrium with and bathed in non-oriented molecules of the same protein. Partially formed spindle fibrils and ribosome-like particles were observed in the mixoplasm when the nuclear envelope had only small discontinuities. Remnants of the envelope are visible throughout division and are probably incorporated into the new envelope in the telophase. Ribosome-like particles are numerous in the metaphase and anaphase spindle but are not seen in the telophase nucleus, once the envelope is reestablished, or in the interphase nucleus.


Author(s):  
Aline Byrnes ◽  
Elsa E. Ramos ◽  
Minoru Suzuki ◽  
E.D. Mayfield

Renal hypertrophy was induced in 100 g male rats by the injection of 250 mg folic acid (FA) dissolved in 0.3 M NaHCO3/kg body weight (i.v.). Preliminary studies of the biochemical alterations in ribonucleic acid (RNA) metabolism of the renal tissue have been reported recently (1). They are: RNA content and concentration, orotic acid-c14 incorporation into RNA and acid soluble nucleotide pool, intracellular localization of the newly synthesized RNA, and the specific activity of enzymes of the de novo pyrimidine biosynthesis pathway. The present report describes the light and electron microscopic observations in these animals. For light microscopy, kidney slices were fixed in formalin, embedded, sectioned, and stained with H & E and PAS.


Author(s):  
K. S. McCarty ◽  
R. F. Weave ◽  
L. Kemper ◽  
F. S. Vogel

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.


Author(s):  
A.J. Mia ◽  
L.X. Oakford ◽  
T. Yorio

Protein kinase C (PKC) isozymes, when activated, are translocated to particulate membrane fractions for transport to the apical membrane surface in a variety of cell types. Evidence of PKC translocation was demonstrated in human megakaryoblastic leukemic cells, and in cardiac myocytes and fibroblasts, using FTTC immunofluorescent antibody labeling techniques. Recently, we reported immunogold localizations of PKC subtypes I and II in toad urinary bladder epithelia, following 60 min stimulation with Mezerein (MZ), a PKC activator, or antidiuretic hormone (ADH). Localization of isozyme subtypes I and n was carried out in separate grids using specific monoclonal antibodies with subsequent labeling with 20nm protein A-gold probes. Each PKC subtype was found to be distributed singularly and in discrete isolated patches in the cytosol as well as in the apical membrane domains. To determine if the PKC isozymes co-localized within the cell, a double immunogold labeling technique using single grids was utilized.


Author(s):  
M. John Hicks ◽  
Leon M. Silverstone ◽  
David G. Gantt ◽  
Catherine M. Flaitz

Although fluoride levels become elevated in sound enamel following a topical fluoride treatment, the caries-preventive effect of fluoride is thought to be due primarily to the role of fluoride in remineralization of clinically undetectable enamel lesions and hypomineralized enamel. During lesion formation, redistribution of fluoride from the enamel surface to the subsurface demineralized enamel occurs. This results in a surface zone with a relatively low fluoride content. In order to maintain an intact surface zone over a carious lesion, it may be necessary to replenish the fluoride levels with an exogenous fluoride source. By acid-etching the lesion surface, a more reactive surface is made available for fluoride interaction. In addition, porosities and etching patterns may be created, allowing for bonding of a caries-resistant resin material to the lesion surface. The purpose of this study was to determine the integrity of the caries-like lesion surface following acid-etching and subsequent stannous fluoride treatment (SnF2).


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