scholarly journals Evidence for an intramembrane component associated with a cellulose microfibril-synthesizing complex in higher plants.

1980 ◽  
Vol 84 (2) ◽  
pp. 315-326 ◽  
Author(s):  
S C Mueller ◽  
R M Brown
Keyword(s):  
Zea Mays ◽  
Plasma Membrane ◽  
Cellulose Microfibril ◽  
Higher Plants ◽  
Freeze Fracture ◽  
Plant Cells ◽  
Intramembrane Particle ◽  
Untreated Plant ◽  
Terminal Complexes ◽  
Particle Rosettes

Freeze-fracture of rapidly frozen, untreated plant cells reveals terminal complexes on E-fracture faces and intramembrane particle rosettes on P-fracture faces. Terminal complexes and rosettes are associated with the ends of individual microfibril impressions on the plasma membrane. In addition, terminal complexes and rosettes are associated with the impressions of new orientations of microfibrils. These structures are sparse within pit fields where few microfibril impressions are observed, but are abundant over adjacent impressions of microfibrils. It is proposed that intramembrane rosettes function in association with terminal complexes to synthesize microfibrils. The presence of a cellulosic microfibril system in Zea mays root segments is confirmed by degradation experiments with Trichoderma cellulase.

Protein secretion in Tetrahymena thermophila: characterization of the secretory mutant strain SB281

1985 ◽  
Vol 78 (1) ◽  
pp. 49-65 ◽  
Author(s):  
N.J. Maihle ◽  
B.H. Satir
Keyword(s):  
Plasma Membrane ◽  
Mutant Strain ◽  
Protein Secretion ◽  
Tetrahymena Thermophila ◽  
Freeze Fracture ◽  
Size Class ◽  
Secretory Product ◽  
Wild Type ◽  
Intramembrane Particle ◽  
Mutant Cells

The ciliated protozoon Tetrahymena thermophila contains membrane-bounded secretory organelles termed mucocysts, the release of which has previously been characterized ultrastructurally as a model system for the events occurring during membrane fusion and protein secretion. Recently, a series of secretory mutant strains of Tetrahymena has been isolated following mutagenesis of a parental wild-type strain designated SB210. In this study, the correlates of non-release in one unique mutant strain of this series, designated SB281, are described. SB281 appears to express a diminished (undetectable) level of the major 34000 Mr proteinaceous secretory product of Tetrahymena, as determined by Western immunoblot analysis and indirect immunofluorescence labelling. Thin-section electron-microscopic studies of these cells reveal that they possess no docked or free mature mucocysts. In addition, freeze-fracture electron microscopy demonstrates that an intramembrane particle array termed the rosette, present in the plasma membrane of wild-type cells above sites of docked mucocysts, is absent in the plasma membrane of mutant SB281 cells. A morphometric analysis of intramembrane particles in the plasma membrane of both wild-type and mutant cells indicates that both strains have a similar intramembrane particle density in both leaflets of the the plasma membrane. Although assembled rosettes are missing in the plasma membrane of mutant cells, a 15 nm intramembrane particle size class does exist in the plasma membrane of the mutant, but this size class is significantly reduced in number relative to wild-type.

scholarly journals Fusion of liposomes with the plasma membrane of epithelial cells: fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections.

1988 ◽  
Vol 107 (6) ◽  
pp. 2511-2521 ◽  
Author(s):  
G Knoll ◽  
K N Burger ◽  
R Bron ◽  
G van Meer ◽  
A J Verkleij
Keyword(s):  
Plasma Membrane ◽  
Apical Cell ◽  
Freeze Fracture ◽  
Freeze Substitution ◽  
Morphological Evidence ◽  
Epithelial Cell Line ◽  
Rapid Freezing ◽  
Intramembrane Particle ◽  
Local Point ◽  
Basolateral Plasma Membrane

The fusion of liposomes with the plasma membrane of influenza virus-infected monolayers of an epithelial cell line, Madin-Darby canine kidney cells (van Meer et al., 1985. Biochemistry. 24:3593-3602), has been analyzed by morphological techniques. The distribution of liposomal lipids over the apical and basolateral plasma membrane domains after fusion was assessed by autoradiography of liposomal [3H]dipalmitoylphosphatidylcholine after rapid freezing or chemical fixation and further processing by freeze substitution and low temperature embedding. Before fusion, radioactivity was solely detected on the apical cell surface, indicating the absence of redistribution artifacts and demonstrating the reliability of lipid autoradiography on both a light and electron microscopical level. After induction of fusion by a low pH treatment, the basolateral plasma membrane domain became progressively labeled, indicative of rapid lateral diffusion of [3H]dipalmitoylphosphatidylcholine in the plasma membrane. Analysis of individual fusion events by freeze fracture after rapid freezing confirmed the rapid diffusion of the liposomal lipids into the plasma membrane, as intramembrane particle-free lipid patches were never observed. After the induction of liposome-cell fusion, well-defined intramembrane particles were present on the otherwise smooth liposomal fracture faces and on the fracture faces of the plasma membrane. Morphological evidence thus was obtained in favor of a local point fusion mechanism with an intramembrane particle as a specific structural fusion intermediate.

Freeze fracture analysis of cellulose terminal synthesizing complexes in Vaucheria

1989 ◽  
Vol 47 ◽  
pp. 772-773
Author(s):  
Shun Mizuta ◽  
R. Malcolm Brown
Keyword(s):  
Single Cells ◽  
Freeze Fracture ◽  
Analytical Techniques ◽  
Sodium Dichromate ◽  
Linear Type ◽  
Terminal Complexes ◽  
Vaucheria Hamata ◽  
Particle Rosettes ◽  
Tinopal Lpw ◽  
Enzyme Complexes

It is well known that cellulose is assembled by specialized enzyme complexes (known as terminal complexes = TCs) located on or within the plasma membrane.At least three types of TCs have been described: a single linear row of particles observed in the brown alga Pelvetia; linear multiple rows in Oocystis and in some green algae; and, hexagonally arranged particle “rosettes” in some algae and solitary rosettes in all land plants. A new linear type of TC has been found in a freshwater xanthophycean alga Vaucheria hamata. Fine structure of the TC has been analyzed by high resolution analytical techniques for freeze-replication. Further examination for its characteristics was carried out by the treatment with inhibitors for cellulose formation(2,6-dichlorobenzonitrile, DCB) and its crystallization (Tinopal LPW).v. hamata was cultured under appropriate light conditions and single cells were cut into small pieces and recultured for two days. They were plasmolyzed in 0.2M NaCl, washed with fresh medium, and then recultured with or without inhibitors for 4-10 h. The cells were placed on Balzers double replica supports and quickly frozen in liquid propane. The supports were inserted in a Balzers double replica holder cooled by liquid nitrogen and placed in the chamber of a Balzers BA 360. Cells were fractured at 2x10-4 Pa and etched for 1 min. Most of the specimens were rotary shadowed (1 rps) with Pt-C at an angle of 30° and coated with carbon. Replicas were cleaned with 50% sulfuric acid containing 5% sodium dichromate and distilled water, and observed at 80Kv on a Philips EM 420.

Cellulose microfibril orientation in Oocystis solitaria: proof that microtubules control the alignment of the terminal complexes

1986 ◽  
Vol 83 (1) ◽  
pp. 223-234
Author(s):  
H. Quader
Keyword(s):  
Cellulose Microfibril ◽  
Drug Application ◽  
Freeze Fracture ◽  
Cellulose Microfibrils ◽  
Cortical Microtubules ◽  
Terminal Complex ◽  
Experimental Proof ◽  
Cellulose Deposition ◽  
Cellulose Microfibril Orientation ◽  
Terminal Complexes

In the green alga Oocystis solitaria microtubules control the regular deposition of cellulose microfibrils. Although it has frequently been suggested that the influence of the cortical microtubules is mediated through the alignment of structures in the plasma membrane, e.g. the cellulose-synthesizing enzymes, experimental proof is lacking. In Oocystis the putative cellulose-synthesizing units, the so-called terminal complexes, can be visualized following freeze-fracture. With respect to the synthesis of a given layer of microfibrils two distinct situations are observable: terminal complex doublets occur before the start of cellulose formation, but are subsequently separated into single terminal complexes by pressure exerted by the crystallizing microfibrils. In order to investigate the effect of anti-microtubular substances on the orientation of the terminal complexes, the state of cellulose deposition at the time of drug application was marked by short (15–30 min) treatment with Congo Red, which causes a morphological change in the terminal complexes. The characteristic alignment of the terminal complexes, both doublets and fragmented single ones, is severely disturbed in cells treated with the herbicide amiprophosmethyl, which is known to interfere with the action of microtubules. The results provide strong evidence that microtubules control the alignment of the putative cellulose-forming units in Oocystis. The observed pattern of interference indicates that the microtubules most probably achieve their control by imposing fluidity channels on the membrane and not via direct links with the terminal complexes.

scholarly journals Immunogold-labelling localization of chlorophyllase at different developmental stages of Pachira macrocarpa leaves

2021 ◽  
Author(s):  
Tzan-Chain Lee ◽  
Kuan-Hung Lin ◽  
Chang-Chang Chen ◽  
Tin-Han Shih ◽  
Meng-Yuan Huang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Thylakoid Membrane ◽  
Developmental Stages ◽  
Higher Plants ◽  
Plant Cells ◽  
Immunogold Labelling ◽  
Transmission Electron

Abstract Background: Chlorophyllases (Chlases) are housekeeping proteins in plant cells. The dephytylating enzymes can catalyze chlorophyll (Chl) to form chlorophyllide, but the distribution of Chlases in plant cells is still an interesting debate. In this study, antibody of PmCLH2 was made and used by immunogold-labelling technique to detect the location of Chlase of Pachira macrocarpa (Pm) leaves at four developmental stages, including young, mature, yellowing, and senesced stages. Results: The transmission electron microscopy results show that Chlases were comprehensively found in portions of chloroplast, such as the inner membrane of the envelope, grana, and the thylakoid membrane of the chloroplast, cytosol, and vacuoles at young, mature, and yellowing stages of Pm leaves, but not in the cell wall, plasma membrane, mitochondria, and nucleus. Conclusions: PmChlases were mainly detected in vacuoles at the senescent stage, but a few were found in the chloroplasts. A pathway is proposed to explain the birth and death of Chl, Chlase, and chloroplasts in higher plants.

scholarly journals Hydrogen evolution and absorption phenomena in plasma membrane of higher plants

2020 ◽  
Author(s):  
Xin Zhang ◽  
Zhao Zhang ◽  
Zehua Su ◽  
Qinghui Zhao ◽  
Tyler W. LeBaron ◽  
...  
Keyword(s):  
Gas Chromatography ◽  
Plasma Membrane ◽  
Abiotic Stress ◽  
Molecular Hydrogen ◽  
Higher Plants ◽  
Plant Cells ◽  
Primary Target ◽  
Non Invasive ◽  
Test Technology ◽  
Uptake Activity

AbstractMolecular hydrogen (H2) exhibits a wide breadth of botanical effects especially in improving the resistance to abiotic stress. However, the primary target is not clear. Bidirectional hydrogenases have been well studied in microbes and alga, but there is a paucity of research in higher plants. Here, we used real-time spectrophotometry, gas chromatography and H2 sensor to detect the H2-evolving and H2-uptake activity of the plasma membrane (PM) vesicles of higher plants. Net H+, Na+ and K+ fluxes were measured using Non-invasive Micro-test Technology (NMT). We found that the PM vesicles of Vigna radiata hypocotyls and Capsicum annuum stems have the ability of both oxidizing and producing H2. This two-way enzymatic activity coupled with the redox of ferricyanide/ferrocyanide and NADH/NAD+. H2 affected the H+ and Na+ fluxes on the surface of V. radiata hypocotyls. These results demonstrate that the PM vesicles of higher plants have the abilities of H2 evolution and absorption. The bidirectional activities may act as a valve to regulate the energy balance in plant cells.

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.

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