Fate of the plasma membrane of Saccharomyces cerevisiae during cell rupture

1973 ◽  
Vol 19 (2) ◽  
pp. 285-290 ◽  
Author(s):  
J. Dubé ◽  
G. Setterfield ◽  
G. Kiss ◽  
C. V. Lusena
Keyword(s):  
Plasma Membrane ◽  
Polyacrylamide Gel Electrophoresis ◽  
Outer Wall ◽  
Yeast Cells ◽  
Wall Material ◽  
Physical Damage ◽  
Intact Cells ◽  
Thin Sections ◽  
Cytoplasmic Membranes ◽  
Mitochondrial Origin

Intact and homogenized yeast cells were studied in thin sections in the electron microscope to determine the fate of the plasma membrane during fractionation. Intact cells possess a unit-membrane plasma membrane closely appressed to the cell wall. After even slight physical damage following limited homogenization in dilute buffer, the plasma membrane collapses away from the wall while the intra-cytoplasmic membranes (ER, vacuolar, nuclear, mitochondrial) dilate and vesiculate. With prolonged homogenization, the plasma membrane fragments and vesiculates and becomes indistinguishable from the remains of the other membranes. Washed wall fractions consist of wall fragments with entrapped vesicles derived from all cellular membranes. Exhaustive digestion of the wall fraction with snail gut enzymes liberates some of the trapped vesicles and results in an undigested, non-membranous, inner layer of wall partially contaminated with outer wall material. Polyacrylamide gel electrophoresis indicates that proteins of the wall "membranes" are partially of mitochondrial origin.

scholarly journals A CYTOCHEMICAL LOCALIZATION OF REDUCTIVE SITES IN A GRAM-POSITIVE BACTERIUM

1964 ◽  
Vol 20 (3) ◽  
pp. 361-375 ◽  
Author(s):  
Woutera van Iterson ◽  
W. Leene
Keyword(s):  
Plasma Membrane ◽  
Cell Periphery ◽  
Gram Positive ◽  
Cytochemical Localization ◽  
Potassium Tellurite ◽  
Thin Sections ◽  
Gram Positive Bacteria ◽  
Cytoplasmic Membranes ◽  
Reducing Activity ◽  
The One

In bacteria the exact location of a respiratory enzyme system comparable to that of the mitochondria of other cells has remained uncertain. On the one hand, the existence of particulate "bacterial mitochondria" has been advocated (Mudd); on the other hand, important enzymes of the respiratory chain were recovered in the cytoplasmic membranes associated with some granular material (Weibull). In order to gain insight into this question, sites of reducing activity were localized in thin sections of bacteria using the reduction of potassium tellurite as an indicator. When this salt was added to the culture medium of Bacillus subtilis, it turned out that in this Gram-positive organism the reduced product is strictly bound at two sites, and that the plasma membrane does not materially gain in electron opacity through deposition of the reduced product. The reduction product is found on or in the membranes of particular organelles, which may possibly be regarded as the mitochondrial equivalents in Gram-positive bacteria, and which are sometimes seen connected to the plasma membrane. The second location is in thin rod-like elements at the cell periphery, possibly the sites from which the flagella emerge.

Cell wall structures of Epicoccum nigrum (Hyphomycetes)

1973 ◽  
Vol 51 (5) ◽  
pp. 1071-1073 ◽  
Author(s):  
J. A. Brushaber ◽  
R. H. Haskins
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Outer Layer ◽  
Secondary Wall ◽  
Outer Wall ◽  
Wall Layer ◽  
Wall Material ◽  
Primary Wall ◽  
Electron Dense Material ◽  
Wall Structures

Two structurally distinct types of secondary wall layers are present in older hyphae in addition to the primary wall. A coarsely fibrous outer wall layer often becomes quite massive and frequently fuses with the outer wall layers of adjacent cells in the formation of hyphal strands. The uneven deposition of this outer layer often produces large verrucosities. The inner secondary wall layer is relatively electron transparent and contains a reticulum of electron-dense lines. The interface of the inner secondary wall with the cytoplasm is often very irregular, and sections of the plasma membrane are frequently overlain by wall material. The outer secondary wall of conidia is composed of an electron-dense material different from that of the outer wall of hyphae. Cells in the multicellular conidia tend to be polyhedral in shape with either very thick primary walls or thin primary walls having a thick inner wall deposit.

Ultrastructure of yeast plasma membrane revealed by a new high-resolution freeze-replica method

1987 ◽  
Vol 45 ◽  
pp. 964-965
Author(s):  
Tadashi Hirano ◽  
Akira Tanaka
Keyword(s):  
Plasma Membrane ◽  
High Resolution ◽  
Single Unit ◽  
Freeze Fracture ◽  
Fracture Morphology ◽  
Replica Method ◽  
Yeast Cells ◽  
Thin Sections ◽  
Yeast Protoplasts ◽  
Fracture Theory

The Freeze-fracture morphology of the plasma membrane and surface of yeast protoplasts has been investigated by a new high resolution freeze-replica method (Tanaka et al.1978). According to freeze-fracture theory, it is generally argued that the plane of cleavage breaks down into the bilayer and then follows the plane of the membrane between the two halves of the lipid. However, when we observed thin sections of replica film of the surface of the freeze-fractured face of intact yeast cells, the single unit membrane was clearly visible between the replica film and the cytoplasm (Fig. 1). Accordingly, in the case of yeast cells, we assume that the plane of cleavage breaks down between the plasma membrane and cell wall.On the other hand, Walzthöng et al. (1982) have shown that surface granules are an artifact or form of contamination produced under the conditions used for the ordinary freeze-replica method employing metal shadowing film.

scholarly journals The degradation and turnover of fucosylated glycoproteins in the plasma membrane of a neuroblastoma-cell line

1977 ◽  
Vol 166 (2) ◽  
pp. 217-223 ◽  
Author(s):  
James E. Hudson ◽  
Terry C. Johnson
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Polyacrylamide Gel Electrophoresis ◽  
Specific Radioactivity ◽  
Complete Medium ◽  
Intact Cells ◽  
Membrane Components ◽  
Turn Over ◽  
Glycoprotein Degradation ◽  
Dual Label

When monolayer cultures of neuroblastoma N2a cells were prelabelled with [3H]fucose to steady state, and then reincubated in complete medium in the presence of unlabelled 40mm-l-fucose, there was a rapid metabolism of fucosylated cellular macromolecules and the specific radioactivity of the acid-insoluble material decreased by 22% within 2h. After this period of time the remaining radioactive glycoproteins appeared to be more stable and the rate of loss of specific radioactivity markedly decreased. Since fucose is known to be associated predominantly with plasma-membrane components, the analysis of fucosylated glycoproteins was characterized in plasma-membrane fractions by polyacrylamide-gel electrophoresis. Two experimental approaches were used to measure glycoprotein degradation and turnover in the cell-surface membranes. In one set of experiments, with a similar incubation procedure to that used with intact cells, three membrane components were rapidly degraded (150000, 130000 and 48000 daltons), but another surface glycoprotein (68000 daltons) appeared to be more slowly metabolized than the mean rate of glycoprotein degradation. The relationship of the degradation of membrane glycoproteins to their turnover was analysed by dual-label experiments that used both [14C]fucose and [3H]fucose. Glycoproteins of the surface membrane of neuroblastoma cells were found to turn over at heterogeneous rates. The components mentioned above that exhibited significantly rapid rates of degradation, were also shown to turn over more rapidly than the average surface component. In addition to the membrane components detected by the use of only [3H]fucose, dual-label experiments illustrated that numerous surface glycoproteins were metabolized more rapidly or slowly than most of the cell-surface constituents.

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

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).

Deformation of Yeast Plasma Membrane by Ethidium Bromide

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.

scholarly journals New methods for confocal imaging of infection threads in crop and model legumes

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Angus E. Rae ◽  
Vivien Rolland ◽  
Rosemary G. White ◽  
Ulrike Mathesius
Keyword(s):  
Cell Wall ◽  
Root Hair ◽  
Periodic Acid ◽  
Confocal Imaging ◽  
Wall Material ◽  
Future Research ◽  
Thin Sections ◽  
New Methods ◽  
Infection Threads ◽  
Imaging Of Infection

Abstract Background The formation of infection threads in the symbiotic infection of rhizobacteria in legumes is a unique, fascinating, and poorly understood process. Infection threads are tubes of cell wall material that transport rhizobacteria from root hair cells to developing nodules in host roots. They form in a type of reverse tip-growth from an inversion of the root hair cell wall, but the mechanism driving this growth is unknown, and the composition of the thread wall remains unclear. High resolution, 3-dimensional imaging of infection threads, and cell wall component specific labelling, would greatly aid in our understanding of the nature and development of these structures. To date, such imaging has not been done, with infection threads typically imaged by GFP-tagged rhizobia within them, or histochemically in thin sections. Results We have developed new methods of imaging infection threads using novel and traditional cell wall fluorescent labels, and laser confocal scanning microscopy. We applied a new Periodic Acid Schiff (PAS) stain using rhodamine-123 to the labelling of whole cleared infected roots of Medicago truncatula; which allowed for imaging of infection threads in greater 3D detail than had previously been achieved. By the combination of the above method and a calcofluor-white counter-stain, we also succeeded in labelling infection threads and plant cell walls separately, and have potentially discovered a way in which the infection thread matrix can be visualized. Conclusions Our methods have made the imaging and study of infection threads more effective and informative, and present exciting new opportunities for future research in the area.

scholarly journals FINE STRUCTURE OF THE GRAM-NEGATIVE BACTERIUM ACETOBACTER SUBOXYDANS

1964 ◽  
Vol 20 (2) ◽  
pp. 217-233 ◽  
Author(s):  
G. W. Claus ◽  
L. E. Roth
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Cell Walls ◽  
Negative Staining ◽  
Homogeneous Layer ◽  
Physical Treatment ◽  
Thin Sections ◽  
Gram Negative ◽  
Acetobacter Suboxydans ◽  
Negative Cell

The morphological features of the cell wall, plasma membrane, protoplasmic constituents, and flagella of Acetobacter suboxydans (ATCC 621) were studied by thin sectioning and negative staining. Thin sections of the cell wall demonstrate an outer membrane and an inner, more homogeneous layer. These observations are consistent with those of isolated, gram-negative cell-wall ghosts and the chemical analyses of gram-negative cell walls. Certain functional attributes of the cell-wall inner layer and the structural comparisons of gram-negative and gram-positive cell walls are considered. The plasma membrane is similar in appearance to the membrane of the cell wall and is occasionally found to be folded into the cytoplasm. Certain features of the protoplasm are described and discussed, including the diffuse states of the chromatinic material that appear to be correlated with the length of the cell and a polar differentiation in the area of expected flagellar attachment. Although the flagella appear hollow in thin sections, negative staining of isolated flagella does not substantiate this finding. Severe physical treatment occasionally produces a localized penetration into the central region of the flagellum, the diameter of which is much smaller then that expected from sections. A possible explanation of this apparent discrepancy is discussed.

scholarly journals Septin-dependent compartmentalization of the endoplasmic reticulum during yeast polarized growth

2005 ◽  
Vol 169 (6) ◽  
pp. 897-908 ◽  
Author(s):  
Cosima Luedeke ◽  
Stéphanie Buvelot Frei ◽  
Ivo Sbalzarini ◽  
Heinz Schwarz ◽  
Anne Spang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Membrane Proteins ◽  
Diffusion Barriers ◽  
Yeast Cells ◽  
Dependent Manner ◽  
Protein Diffusion ◽  
Ultrastructural Studies ◽  
Bud Neck ◽  
Er Membrane

Polarized cells frequently use diffusion barriers to separate plasma membrane domains. It is unknown whether diffusion barriers also compartmentalize intracellular organelles. We used photobleaching techniques to characterize protein diffusion in the yeast endoplasmic reticulum (ER). Although a soluble protein diffused rapidly throughout the ER lumen, diffusion of ER membrane proteins was restricted at the bud neck. Ultrastructural studies and fluorescence microscopy revealed the presence of a ring of smooth ER at the bud neck. This ER domain and the restriction of diffusion for ER membrane proteins through the bud neck depended on septin function. The membrane-associated protein Bud6 localized to the bud neck in a septin-dependent manner and was required to restrict the diffusion of ER membrane proteins. Our results indicate that Bud6 acts downstream of septins to assemble a fence in the ER membrane at the bud neck. Thus, in polarized yeast cells, diffusion barriers compartmentalize the ER and the plasma membrane along parallel lines.

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