Mitochondria attached to desmosomes in the ciliary epithelia of human,monkey, and rabbit eyes

In the normal ciliary epithelia of the rhesus monkey, owl monkey, albino rabbit, and human eye, a previously unreported relationship exists between mitochondria and certain desmosomes. At these sites, two mitochondria appear like "sentinels" attached to the cytoplasmic surfaces of their respective sides of a desmosome. In other instances, only one side of the junction may be afforded an associated mitochondrion. In each case the cytoplasmic filaments of the desmosome are seen to blend with the outer membrane of the mitochondrion. The relationship between desmosomes and mitochondria in the ciliary epithelium is unique among ocular tissues. A survey of ocular epithelia in the various species examined, failed to give any evidence of similar junctional/organelle complexes. Various functional roles for this relationship are discussed including the possibility that the mitochondria could control the cytoplasmic calcium ion concentration in the microenvironment of their associated desmosomal junctions.     

Cytoplasmic Ultrastructural Changes During Microsporogenesis of Gossypium hirsutum:With Emphasis on

Conspicuous cytoplasmic changes took place during the microsporogenesis of Gossypium hirsutum L. These changes mainly involved in the ribosomes, plastids and mitochondria. During meiotic prophase 1, the ribosome population of the cytoplasm diminished and reached to aminimum during pachytene--diplotene interval, and the membrane structures of both plastids and mitochondria turned unclear. In metaphase I, cytoplasmic ribosome population restored to premeiotic level; plastids and mitochondria also regained their normal structures. The disintegration of nucleoloids from nucleus was the main mechanism for the restoration of ribosome population in metaphase I cytoplasm. Endoplasmic reticulum may play an important role in the elimination and protection of part of cytoplasmic ribosomes during prophase I. These obvious cytoplasmic changes are considered to be relevant to sporophyte-gametophyte transition.     

In vitro evidence for the involvement of mitochondrial folate metabolism in the supply of cytoplasmic one-carbon units

We present in vitro evidence for a novel intercompartmental pathway in which folate-mediated reactions in mitochondria generate one-carbon units for utilization in cytoplasmic processes. Rat liver mitochondria are shown to contain the enzymatic activities for catabolism of serine or sarcosine to produce formate. Intact mitochondria rapidly convert the 3-carbon of serine or the N-methyl group of sarcosine to formate, which exits the mitochondria. Labeled formate is incorporated into purine by a cytoplasmic purine synthesizing system only after activation to 10-formyl-THF via the ATP-dependent 10-formyl-THF synthetase reaction. In a coupled system where one-carbon donors are catabolized by mitochondria before addition to the cytoplasmic purine synthesizing system, incorporation into purine shows a marked dependence on ATP. These observations demonstrate that mitochondria can metabolize one-carbon donors via THF-dependent reactions to the level of formate which then exits mitochondria for utilization in the cytoplasm. The proposed pathway is discussed in relation to genetic evidence for its operation in vivo as well as compartmentation of folate coenzymes and their one-carbon units.     

The morphological relationship between mitochondria and cytoplasmic membranes of the follicular oocyte in domestic pig

Summary An ultrastructural study of the mature follicular oocytes in domestic pig demonstrate a morphological relationship between the mitochondria and the cytoplasmic membranes immediately surrounding the yolk globules of the cells. Frequently, the cytoplasmic membranes are observed to be in close proximity of the mitochondria or are found to be continuous with the outer mitochondrial membrane. Sometimes the cytoplasmic membranes are found to display the formation of one or more oval loops of different diameter located at their presumed ends or free in the nearby cytoplasm. The significance of these observations is discussed in the light of the available informations, which suggest that the cytomembrane system in certain phases of development may take part in the formation of mitochondria.This work was supported by the Agricultural Research Council of Norway.     

Yeast mitochondrial and cytoplasmic valyl-tRNA synthetases

Yeast mitochondrial tRNA synthetase has been partially purified and chromatographic, catalytic and antigenic properties have been compared to the cytoplasmic homologous enzyme from yeast. No significant differences could be observed between the two enzymes with respect to their behaviour during ammonium sulfate precipitation or in chromatographic separation on DEAE cellulose, hydroxylapatite and Sephadex G 200. The Km of the two enzymes toward tRNAs from yeast mitochondria, yeast cytoplasm or E. coli are pratically identical. The antigenic properties of the two enzymes are very similar; antisera against either the mitochondria or the cytoplasmic enzyme lead to the inhibition of their catalytic properties. The mitochondrial ValRS is formed by a single polypeptide chain whose molecular weight is 125,000 daltons, a value very close to that of the yeast cytoplasmic enzyme.     

Ribosomal Ribonucleic Acids of Chloroplastic and Mitochondrial Preparations

RNA prepared from fractions of chloroplasts and mitochondria sedimented at rates characteristic of ribosomal RNA. A predominance of the 18S species was frequently observed in preparations from chloroplasts from romaine lettuce and endive. The usual distribution, a preponderance of the 28S species, was observed in studies on tomato and spinach chloroplasts and mitochondria from mushroom and cauliflower. Comparisons of the base composition of RNA from organelles with their cytoplasmic ribosomal counterparts revealed that the 18S component from romaine lettuce chloroplast was different. A marginally significant difference was observed in the 28S particle from mushroom mitochondria preparations whereas distinct differences, reflected in all the bases, were seen when the 18S component of cauliflower mitochondria preparations was compared with cytoplasmic RNA.     

Mitochondrial morphology in Basidiobolus haptosporus

Young hyphal cells of the potentially zoopathogenic fungus Basidiobolus haptosporus characteristically exhibit unusual proportions of annulate views of mitochondria in the two-dimensional perspective of thin sections. Such views exhibit a central space containing cytoplasmic ground substance and often profiles of other cytoplasmic organelles (lipid bodies, other mitochondrial forms, and especially crystalloid-containing microbodies). Three-dimensional projections are presented to suggest that these mitochondria have assumed the form of a goblet-shaped enclosure, and that the various annulate views are the consequence of plane of section viewed by electron microscopy. Their frequent occurrence and consistent morphology argues against their being random expressions of mitochondrial plasticity, but rather for close spatial associations amongst cytoplasmic organelles of young hyphae. When the fungus is grown on xanthine or its catabolites as sole sources of nitrogen, there is a proliferation of crystalloid-containing microbodies, double-membraned vesicles, and ovate to ellipsoidal mitochondria. Annulate views of mitochondria then are no longer observed, but microbodies again frequently appear in close association with mitochondria and at times in intimate contact with the mitochondrial outer membrane.     

RNA biosynthesis in mitochondria under conditions of prolonged inhibition of protein biosynthesis in rat liver cytoplasm]

When cytoplasmic protein synthesis is inhibited by cycloheximide (CHI) in vivo synthesis of water-soluble mitochondrial proteins and of mitochondrial RNA is decreased. These changes measured in isolated rat liver mitochondria are similar to those observed in vivo and correlate with the changes the synthesis of water-soluble proteins in mitochondria. When the cytoplasmic fraction (30,000 g-supernatant) had been added to the mitochondria showing decreased RNA synthesis, the RNA synthesis increased to the control level (the incubation conditions were favourable for the protein transport from microsomes to mitochondria). RNA synthesis in mitochondria was not stimulated by cytoplasmic fractions from the CHI-pretreated rats. After prolonged dialysis these fraction stimulated RNA synthesis even to a greater extent than cytoplasmic fractions from the untreated animals. Mitochondrial RNA polymerase activity (measured in mitochondrial extracts supplemented with exogenous DNA) was higher in extracts of mitochondria from livers of normal rats than in extracts of mitochondria from livers of animals injected with CHI.     

Targeting of tRNA into yeast and human mitochondria: the role of anticodon nucleotides

In vivo, yeast mitochondria import a single cytoplasmic tRNA, tRNA(CUU)Lys, while human mitochondria do not import any cytoplasmic tRNA. We have previously demonstrated that both yeast and human isolated mitochondria can specifically internalize tRNA(CUU)Lys, several of its mutant versions and some mutant versions of yeast cytosolic tRNA(UUU)Lys (not imported in vivo). Aminoacylation of tRNA(CUU)Lys by the cytoplasmic lysyl-tRNA synthetase was a prerequisite for its import. Here we are studying the influence of one-base replacements in the anticodon of tRNAs(Lys) on their aminoacylation, on binding to the precursor of the mitochondrial lysyl-tRNA synthetase (carrier protein directing the import), and on the efficiency of import into isolated yeast and human mitochondria. We show that the base U35 is the main identity element for the yeast cytoplasmic lysyl-tRNA synthetase. The single replacement that abolished import was C34G, while all the others only modulated the import efficiency. The need of aminoacylation for import and for interaction with the carrier protein was shown only for a subset of mutant versions, while the others could be recognized and internalized without aminoacylation or in misacylated forms.     

Exceptional inheritance of plastids via pollen in Nicotiana sylvestris with no detectable paternal mitochondrial DNA in the progeny

Plastids and mitochondria, the DNA‐containing cytoplasmic organelles, are maternally inherited in the majority of angiosperm species. Even in plants with strict maternal inheritance, exceptional paternal transmission of plastids has been observed. Our objective was to detect rare leakage of plastids via pollen in Nicotiana sylvestris and to determine if pollen transmission of plastids results in co‐transmission of paternal mitochondria. As father plants, we used N. sylvestris plants with transgenic, selectable plastids and wild‐type mitochondria. As mother plants, we used N. sylvestris plants with Nicotiana undulata cytoplasm, including the CMS‐92 mitochondria that cause cytoplasmic male sterility (CMS) by homeotic transformation of the stamens. We report here exceptional paternal plastid DNA in approximately 0.002% of N. sylvestris seedlings. However, we did not detect paternal mitochondrial DNA in any of the six plastid‐transmission lines, suggesting independent transmission of the cytoplasmic organelles via pollen. When we used fertile N. sylvestris as mothers, we obtained eight fertile plastid transmission lines, which did not transmit their plastids via pollen at higher frequencies than their fathers. We discuss the implications for transgene containment and plant evolutionary histories inferred from cytoplasmic phylogenies.     

Energetics of Ehrlich ascites mitochondria: membrane potential of isolated mitochondria and mitochondria within digitonin-permeabilized cells

Ehrlich ascites tumour cells were treated with digitonin so that they became permeable for low-molecular-weight compounds but, at certain concentrations of digitonin, retained most of their cytoplasmic proteins. Respiration of mitochondria with exogenous substrates and their membrane potential could thus be measured in situ by means of oxygen electrode and tetraphenylphosphonium-sensitive electrode, respectively. The results were compared with data from similar measurements on mitochondria isolated from such digitonin-permeabilized cells. Isolated mitochondria and mitochondria in situ oxidized succinate at similar rates and developed membrane potential of comparable magnitude. Both preparations also exhibited an identical nonlinear relationship between resting state respiration (titrated with a respiratory inhibitor) and the membrane potential. In the cells permeabilized with low concentrations of digitonin (i.e., retaining most of cytoplasmic proteins) and suspended in medium containing NaCl and other major anions and cations at concentrations close to those in mammalian plasma, anaerobiosis did not produce a decrease in the mitochondrial membrane potential, which was collapsed only after a subsequent addition of oligomycin. In this medium, glucose had little effect on either respiration or the membrane potential.     

Cholinesterase in larvae of the ascidian,Ciona intestinalis,developing from fragments cut from centrifuged eggs

Development Genes and Evolution - UnfertilizedCiona eggs were centrifuged, stratifying their mitochondria and some other cytoplasmic components. Each centrifuged egg had a mitochondria-free,...     

Toxicity of familial ALS-linked SOD1 mutants from selective recruitment to spinal mitochondria

One cause of amyotrophic lateral sclerosis (ALS) is mutation in ubiquitously expressed copper/zinc superoxide dismutase (SOD1), but the mechanism of toxicity to motor neurons is unknown. Multiple disease-causing mutants, but not wild-type SOD1, are now demonstrated to be recruited to mitochondria, but only in affected tissues. This is independent of the copper chaperone for SOD1 and dismutase activity. Highly preferential association with spinal cord mitochondria is seen in human ALS for a mutant SOD1 that accumulates only to trace cytoplasmic levels. Despite variable proportions that are successfully imported, nearly constant amounts of SOD1 mutants and covalently damaged adducts of them accumulate as apparent import intermediates and/or are tightly aggregated or crosslinked onto integral membrane components on the cytoplasmic face of those mitochondria. These findings implicate damage from action of spinal cord-specific factors that recruit mutant SOD1 to spinal mitochondria as the basis for their selective toxicity in ALS.     

Nucleo-cytoplasmic interactions affect RNA editing of cox2, atp6 and atp9 in alloplasmic male-sterile rice (Oryza sativa L.) lines

RNA editing plays an important role in the regulation of mitochondrial gene expression in flowering plants. In this study, we examined RNA editing of the mitochondrial genes cox2, atp6 and atp9 in five isonuclear alloplasmic male-sterile lines (IAMSLs) of rice to investigate whether different cytoplasmic types affect RNA editing. Although many editing sites were conserved among the three genes, we found that the editing efficiency of certain sites was significantly different between different IAMSLs or between IAMSLs and their corresponding cytoplasmic donor CMS lines. Furthermore, several editing sites were found to be either present or absent in certain IAMSLs and their corresponding CMS lines. These results indicate that nuclear loci, as well as unknown editing factors within the mitochondria of different cytoplasmic types, may be involved in RNA editing, and they suggest that RNA editing in plant mitochondria is affected by nucleo-cytoplasmic interactions.     

Changes of Plastids Mitochondria and Their DNA in the Egg Cell Before and After Fertilization in Pharbitis

The ultrastructure and cytoplasmic DNA in the egg cell and zygote of Pharbitis purpurea, (L.) Voyght and P. limbata Lindl. which were studied with electron microscopy and DNA epifiuorescence microscopy. The egg cell before fertilization was highly vacuolated with only a few cytoplasmic plastids and mitochondria. Plastids were spherical and/or rod- shaped containing 1 ～ 2 large starch grains. Most of the mitochondria were cup and/or circular. The cytoplasm in the zygote was much more abundant than that in the egg cell. The number of plastids and their electronic density were greatly increased, in most of which containing osmiophilic bodies. The mitochondria were rich and spherical-shaped in the zygote. Two types of cytoplasmic DNA nucleoids were detected in the egg cell, the more abundant one being big and circle-shaped and the other dot-shaped. Only dot-shaped nucleoids were present in the zygote. The content of nucleoids in the zygote was much less than that in the egg cell. Authors propose that some cytoplasmic DNA may degenerate after fertilization. The ultrastructural characteristics of the egg cell and the reduction of cytoplasmic DNA in the zygote may related to the mechanisms of plastid unipaternal inheritance in Pharbitis.     

The products of mitochondria-bound cytoplasmic polysomes in yeast

Experiments were undertaken to examine the fate and composition of polypeptides synthesized on cytoplasmic polysomes associated with the outer mitochondrial membrane of Saccharomyces cerevisiae. Mitochondria with their associated cytoplasmic polysomes were isolated from growing yeast spheroplasts and placed in a polypeptide chain completion system together with [35S]methionine. Of the total products synthesized in the readout system, 80 to 85% remain associated with the mitochondria after sucrose gradient centrifugation. Most of the labeled products are resistant to papain digestion unless the membranes are disrupted by treatment with detergent or shaking with glass beads. When free cytoplasmic polysomes were translated in the presence of [35S]methionine and incubated with mitochondria, only about 20% of the labeled polypeptides remain associated with the mitochondria; furthermore, most of these products are equally sensitive to papain digestion in the presence or absence of detergent. These results support the view that the cytoplasmic polysomes associated with the outer mitochondrial membrane of yeast facilitate the segregation of newly synthesized proteins into the organelle. The proportion of the alpha, beta, and gamma subunits of the F1-ATPase was determined among the products synthesized by mitochondria-bound and free cytoplasmic polysomes. By double antibody precipitation and immunoreplicate electrophoresis, we find that the proportion of the subunits of F1-ATPase is much greater among the products of the mitochondria-bound polysomes than those synthesized on free polysomes.     

Distribution of mitochondria within Muller cells – II. Post-natal development of the rabbit retinal periphery in vivo and in vitro: dependence on oxygen supply

The occurrence and localization of mitochondria within glial (Muller) cells and neurons of the peripheral (avascular) rabbit retina was studied electron microscopically and by immunocytochemical demonstration of the mitochondrial enzyme GABA transaminase (GABA-T). Post-natal development in vivo was compared with development of organ cultures from neonatal rabbit retinae, grown over 2 weeks in vitro. The adult pattern of mitochondrial localization (restriction to the sclerad end of the cells) was observed from the beginning of enzyme expression at early post-natal stages. However, when neonatal retinal pieces were grown in vitro with their vitread surface exposed to the air, their Muller cells contained mitochondria along most of their length. When functionally developed retinae from postnatal day 14 were explanted in vitro, they retained their sclerad mitochondrial distribution for almost 24 h but thereafter the inner portions of their cytoplasm became occupied by mitochondria within a few hours. This was achieved mainly by mitochondrial migration rather than by formation of new mitochondria because it was not prevented by cycloheximide-induced inhibition of protein synthesis. These data support the following hypotheses: (1) the mitochondrial distribution in Muller cells is determined by the local cytoplasmic O2 pressure (pO2), (2) existing mitochondria move towards cytoplasmic regions of sufficient pO2 by rather rapid migration and (3) the start of this migration is delayed by almost 24 h due to the action of as yet unknown control mechanisms. In contrast, the mitochondrial content of retinal ganglion and amacrine cells in the vitread retinal layers was virtually independent of the source and level of oxygen supply.     

Regulation of thyroid hormones of the interaction of mitochondria with low-molecular weight cytoplasmic mediators, induced by phosphate- dependent transport of K+ and H+ ions through the mitochondrial inner membrane

In vivo thyroid hormones control the binding to mitochondria of low molecular weight water-soluble cytoplasmic mediators that are capable to induce oxidative phosphorylation uncoupling, by increasing the sensitivity of mitochondria to the effects of these mediators. In hyperthyroid rat liver mitochondria cytoplasmic mediators stimulate the phosphate-dependent transport of K+ and H+ in a greater degree than in liver mitochondria of control rats. The increase in the oxidative phosphorylation uncoupling by cytoplasmic mediators is one of mechanisms of thermogenesis stimulation by thyroid hormones.     

CYTOLOGICAL DIFFERENCES BETWEEN A CYTOPLASMIC MALE STERILE TOBACCO CYBRID AND ITS FERTILE COUNTERPART DURING EARLY ANTHER DEVELOPMENT

Ultrastructural differences were detected between a cytoplasmic male sterile tobacco cybrid (Nicotiana sp.) formed by protoplast fusion and normal, fertile tobacco. Cell structure was compared between anther primordia from normal, fertile tobacco and anther primordia from the cybrid using stereological methods. Particular emphasis was placed on the ultrastructure of mitochondria because of their known relationship to cytoplasmic male sterility in this cybrid and other plants. The volume density of mitochondria in cybrid anther primordia (6.3%) was significantly lower than in normal, fertile anther primordia (10.1%), although mitochondria from both plants contained similar amounts of cristae and profiles were of similar relative area. Dictyosomes and vacuoles also differed in volume density but not at a statistically significant level. Although the volume density of plastids did not differ, a larger amount of starch was stored within plastids in cybrid anther primordia than in normal, fertile anther primordia. These results are compatible with the hypothesis that an abnormally low rate of mitchondrial replication, and the resultant limit on adenosine triphosphate production, could contribute to cytoplasmic male sterility in the cybrid.     

Ultracytochemical evidence for a proton-pump adenosine triphosphatase in chick osteoclasts

Summary The distribution of Mg^{+ +}-ATPase in osteoclasts along the endosteal surface of the chick tibia was investigated by neutral and alkaline pH cytochemical methods at the electron-microscopic level. Reaction product was observed in mitochondria, cytoplasmic vesicles, and ruffled-border membrane. Levamisole, ouabain, and vanadate did not affect the enzymatic activity. Para-chloromercuribenzoic acid (PCMB) prevented staining of mitochondria, ruffled border, and most cytoplasmic vesicles. Tri-n-butyltin decreased the amount of reaction product in cytoplasmic vesicles and ruffled-border membrane, but did not inhibit reaction product formation within mitochondria. Duramycin, which is a potent inhibitor for proton-pump ATPase, blocked reaction-product formation along the ruffled-border membrane, in mitochondria, and in cytoplasmic vesicles at alkaline pH, but not at neutral pH. It is concluded that the alkaline pH method for Mg^{+ +}-ATPase appears to demonstrate sites of proton-pump ATPase activity.