Response of yeast cells to high glucose involves molecular and physiological differences when compared to other osmostress conditions

InSaccharomyces cerevisiae, mitochondrial morphology changes when cells are shifted between nonfermentative and fermentative carbon sources. Here, we show that cells ofS. cerevisiaegrown in different glucose concentrations display different mitochondrial morphologies. The morphology of mitochondria in the cells growing in 0.5% glucose was similar to that of mitochondria in respiring cells. However, the mitochondria of cells growing in higher glucose concentrations (2% and 4%) became fragmented after growth in these media, due to the production of acetic acid; however, the fragmentation was not due to intracellular acidification. From a screen of mutants involved in sensing and utilizing nutrients, cells lackingTOR1had reduced mitochondrial fragmentation, and autophagy was found to be essential for this reduction. Mitochondrial fragmentation in cells grown in high glucose was reversible by transferring them into conditioned medium from a culture grown on 0.5% glucose. Similarly, the chronological lifespan of cells grown in high glucose medium was reduced, and this phenotype could be reversed when cells were transferred to low glucose conditioned medium. These data indicate that chronological lifespan seems correlated with mitochondrial morphology of yeast cells and that both phenotypes can be influenced by factors from conditioned medium of cultures grown in low glucose medium.

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Heterogeneity of Size and Distribution of Intramembrane Particles in the Plasma Membrane of Yeast

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080407 ◽

1977 ◽

Vol 35 ◽

pp. 596-597

Author(s):

E. Keyhani

Keyword(s):

Plasma Membrane ◽

Candida Utilis ◽

Synthetic Medium ◽

Freeze Fracture ◽

Translational Diffusion ◽

Yeast Cells ◽

Distilled Water ◽

Intramembrane Particles ◽

The Matrix ◽

Water Cell

The matrix of biological membranes consists of a lipid bilayer into which proteins or protein aggregates are intercalated. Freeze-fracture techni- ques permit these proteins, perhaps in association with lipids, to be visualized in the hydrophobic regions of the membrane. Thus, numerous intramembrane particles (IMP) have been found on the fracture faces of membranes from a wide variety of cells (1-3). A recognized property of IMP is their tendency to form aggregates in response to changes in experi- mental conditions (4,5), perhaps as a result of translational diffusion through the viscous plane of the membrane. The purpose of this communica- tion is to describe the distribution and size of IMP in the plasma membrane of yeast (Candida utilis).Yeast cells (ATCC 8205) were grown in synthetic medium (6), and then harvested after 16 hours of culture, and washed twice in distilled water. Cell pellets were suspended in growth medium supplemented with 30% glycerol and incubated for 30 minutes at 0°C, centrifuged, and prepared for freeze-fracture, as described earlier (2,3).

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Ultrathin frozen sections of yeast without aldehyde prefixation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081152 ◽

1978 ◽

Vol 36 (2) ◽

pp. 58-59

Author(s):

K. J. Böhm ◽

a. E. Unger

Keyword(s):

Aqueous Solutions ◽

Biological Material ◽

Yeast Cells ◽

Frozen Sections ◽

Good Preservation ◽

Ultrathin Frozen Sections

During the last years it was shown that also by means of cryo-ultra-microtomy a good preservation of substructural details of biological material was possible. However the specimen generally was prefixed in these cases with aldehydes.Preparing ultrathin frozen sections of chemically non-prefixed material commonly was linked up to considerable technical and manual expense and the results were not always satisfying. Furthermore, it seems to be impossible to carry out cytochemical investigations by means of treating sections of unfixed biological material with aqueous solutions.We therefore tried to overcome these difficulties by preparing yeast cells (S. cerevisiae) in the following manner:

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Plasmalemma localisation of inorganic phosphate in B cells pancreas

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100083515 ◽

1978 ◽

Vol 36 (2) ◽

pp. 528-529

Author(s):

F. B. P. Wooding ◽

K. Pedley ◽

N. Freinkel ◽

R. M. C. Dawson

Keyword(s):

Electron Microscope ◽

B Cells ◽

B Cell ◽

Pancreatic Islets ◽

High Glucose ◽

Lead Acetate ◽

Inorganic Phosphate ◽

Slight Modification ◽

Uranyl Acetate ◽

Lead Phosphate

Freinkel et al (1974) demonstrated that isolated perifused rat pancreatic islets reproduceably release up to 50% of their total inorganic phosphate when the concentration of glucose in the perifusion medium is raised.Using a slight modification of the Libanati and Tandler (1969) method for localising inorganic phosphate by fixation-precipitation with glutaraldehyde-lead acetate we can demonstrate there is a significant deposition of lead phosphate (identified by energy dispersive electron microscope microanalysis) at or on the plasmalemma of the B cell of the islets (Fig 1, 3). Islets after incubation in high glucose show very little precipitate at this or any other site (Fig 2). At higher magnification the precipitate seems to be intracellular (Fig 4) but since any use of osmium or uranyl acetate to increase membrane contrast removes the precipitate of lead phosphate it has not been possible to verify this as yet.

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Deformation of Yeast Plasma Membrane by Ethidium Bromide

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103334 ◽

1988 ◽

Vol 46 ◽

pp. 254-255

Author(s):

E. Keyhani

Keyword(s):

Plasma Membrane ◽

Thin Section ◽

Ethidium Bromide ◽

Freeze Fracture ◽

Mutagenic Effect ◽

Yeast Cells ◽

Hydrophobic Region ◽

Thin Section Electron Microscopy ◽

Section Electron ◽

Deep Pits

The mutagenic effect of ethidium bromide on the mitochondrial DNA is well established. Using thin section electron microscopy, it was shown that when yeast cells were grown in the presence of ethidium bromide, besides alterations in the mitochondria, the plasma membrane also showed alterations consisting of 75 to 110 nm-deep pits. Furthermore, ethidium bromide induced an increase in the length and number of endoplasmic reticulum and in the number of intracytoplasmic vesicles.Freeze-fracture, by splitting the hydrophobic region of the membrane, allows the visualization of the surface view of the membrane, and consequently, any alteration induced by ethidium bromide on the membrane can be better examined by this method than by the thin section method.Yeast cells, Candida utilis. were grown in the presence of 35 μM ethidium bromide. Cells were harvested and freeze-fractured according to the procedure previously described.

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Three-Dimensional Immunoelectron Microscopy of Yeast Cells by a New High-Resolution Replica Method

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100119594 ◽

1985 ◽

Vol 43 ◽

pp. 562-563

Author(s):

Hirano T. ◽

M. Yamaguchi ◽

M. Hayashi ◽

Y. Sekiguchi ◽

A. Tanaka

Keyword(s):

High Resolution ◽

Plasma Polymerization ◽

Three Dimensional ◽

Freeze Fracture ◽

Replica Method ◽

Yeast Cells ◽

Dynamic Aspect ◽

Replica Technique ◽

Gaseous Hydrocarbon ◽

Transmission Electron

A plasma polymerization film replica method is a new high resolution replica technique devised by Tanaka et al. in 1978. It has been developed for investigation of the three dimensional ultrastructure in biological or nonbiological specimens with the transmission electron microscope. This method is based on direct observation of the single-stage replica film, which was obtained by directly coating on the specimen surface. A plasma polymerization film was deposited by gaseous hydrocarbon monomer in a glow discharge.The present study further developed the freeze fracture method by means of a plasma polymerization film produces a three dimensional replica of chemically untreated cells and provides a clear evidence of fine structure of the yeast plasma membrane, especially the dynamic aspect of the structure of invagination (Figure 1).

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