Membranes in lupin root nodules. I. The role of Golgi bodies in the biogenesis of infection threads and peribacteroid membranes

1978 ◽  
Vol 30 (1) ◽  
pp. 129-149 ◽  
Author(s):  
J.G. Robertson ◽  
P. Lyttleton ◽  
S. Bullivant ◽  
G.F. Grayston

The process of infection of lupin nodule cells by rhizobia was examined using thin-section and freeze-fracture electron-microscopic techniques to characterize the properties of different membranes and to establish relationships between them. The membranes of the Golgi bodies and the endoplasmic reticulum stained with zinc iodide-osmium tetroxide but not with phosphotungstic acid or silver. By contrast the infection thread membranes, peribacteroid membranes, plasma membranes and membranes of cytoplasmic vesicles did not stain with zinc iodide-osmium tetroxide but stained with phosphotungstic acid and silver. The peribacteroid membranes and plasma membranes are, however, different from each other since the particle density on the E face of freeze-fracture replicas of plasma membranes was twice that on the E face of the peribacteroid membranes. An examination of the tips of the infection threads in the cytoplasm of the plant cells, showed that the rhizobia bud off from the infection threads enclosed in the infection thread membranes. The rhizobia continue to divide still surrounded by membranes of plant origin, namely the peribacteroid membranes. Cytoplasmic vesicles are observed in both thin-section and freeze-fracture preparations of nodule tissue closely associated with, and apparently produced by, Golgi bodies. Formation of the walls and membranes of the infection threads and of the peribacteroid membranes involves fusion of the cytoplasmic vesicles with these membranes. It is proposed that the process of infection of plant cells in lupin nodules involves a change in the function of the Golgi body system for the biogenesis of plant cell walls and plasma membranes to include the synthesis of the walls and membranes of the infection threads and also the peribacteroid membranes.

1980 ◽  
Vol 86 (1) ◽  
pp. 64-76 ◽  
Author(s):  
J P Caulfield ◽  
G Korman ◽  
A E Butterworth ◽  
M Hogan ◽  
J R David

Neutrophils and eosinophils adhering to the surface of schistosomula of Schistosoma mansoni have been partially or completely detached with hypertonic sucrose or by pipetting. The sucrose-treated neutrophils are attached only in areas where there are pentalaminar fusions between the neutrophil and tegumental membranes, suggesting that these fusions attach the cells to the parasites. Pipetting breaks many of the attached cells. In thin section, the tegumental membrane underlying these cells is seen to be pentalaminar. By freeze-fracture techniques, modified attachment areas are found. The edge zone often appears as a single strand of intramembrane particles (IMPs) on the P2 face and as a groove on the E2 face. The edge zone may also have large discontinuities, in which case it no longer separates membrane faces of unequal IMP density from one another. In addition, the IMPs on the IMP-rich areas become aggregated and surrounded by craters in the membrane. These experiments suggest that the fusions may be the mechanism by which the parasite acquires some host membrane components on its surface. On the other hand, eosinophil plasma membranes are seen adhering to a layer of electron-dense material on the parasite after the cells have been disrupted by pipetting. This suggests that eosinophils adhere to the parasite surface through their discharged granule material and not by membrane fusions.


1972 ◽  
Vol 47 (4) ◽  
pp. 195-200 ◽  
Author(s):  
J.-C. Roland ◽  
Carole A. Lembi ◽  
D. James Morré

1979 ◽  
Vol 81 (2) ◽  
pp. 411-425 ◽  
Author(s):  
N Kalderon ◽  
N B Gilula

Myoblast fusion has been studied in cultures of chick embryonic muscle utilizing ultrastructural techniques. The multinucleated muscle cells (myotubes) are generated by the fusion of two plasma membranes from adjacent cells, apparently by forming a single bilayer that is particle-free in freeze-fracture replicas. This single bilayer subsequently collapses, and cytoplasmic continuity is established between the cells. The fusion between the two plasma membranes appears to take place primarily within particle-free domains (probably phospholipid enriched), and cytoplasmic unilamellar, particle-free vesicles are occasionally associated with these regions. These vesicles structurally resemble phospholipid vesicles (liposomes). They are present in normal myoblasts, but they are absent in certain fusion-arrested myoblast popluations, such as those treated with either 5-bromo-deoxyuridine (BUdR), cycloheximide (CHX), or pospholipase C (PLC). The unilamellar, particle-free vesicles are present in close proximity to the plasma membranes, and physical contact is observed frequently between the vesicle membrane and the plasma membrane. The regions of vesicle membrane-plasma membrane interaction are characteristically free of intramembrane particles. A model for myoblast fusion is presented that is based onan interpretation of these observations. This model suggests that the cytoplasmic vesicles initiate the generation of particle-depleted membrane domains, both being essential components in the fusion process.


1974 ◽  
Vol 40 (6) ◽  
pp. 696-705 ◽  
Author(s):  
Eiichi Tani ◽  
Kimiyuki Ikeda ◽  
Susumu Kudo ◽  
Shogo Yamagata ◽  
Noboru Higashi ◽  
...  

✓ The capillaries in two cerebellar hemangioblastomas were studied by thin-section and freeze-fracture techniques. Fenestrae were found in the attenuated portions of the endothelium, and plasmalemmal vesicles in the nonfenestrated portions. In freeze-fracture preparations the fenestrae of the endothelial plasma membrane were about 450 to 550 A in diameter. They appeared as holes and “necks” in clusters of about 40 to 60 per µm2. When the fracture plane passed in a stepwise fashion from the luminal plasma membrane into the contraluminal plasma membrane, the fenestrae at the fracture edge involved both plasma membranes.


Author(s):  
Juan Mora-Galindo ◽  
Jorge Arauz-Contreras

The zinc iodide-osmium tetroxide (ZIO) technique is presently employed to study both, neural and non neural tissues. Precipitates depends on cell types and possibly cell metabol ism as well.Guinea pig cecal mucosa, already known to be composed of epithelium with cells at different maturation stages and lamina propria which i s formed by morphologically and functionally heterogeneous cell population, was studied to determine the pat tern of ZIO impregnation. For this, adult Guinea pg cecal mucosa was fixed with buffered 1.2 5% g 1 utara 1 dehyde before incubation with ZIO for 16 hours, a t 4°C in the dark. Further steps involved a quick sample dehydration in graded ethanols, embedding in Epon 812 and sectioning to observe the unstained material under a phase contrast light microscope (LM) and a transmission electron microscope (TEM).


Author(s):  
E. Keyhani

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.


Biologia ◽  
2008 ◽  
Vol 63 (2) ◽  
Author(s):  
Lucia Mikolajová ◽  
Halina Vargová ◽  
Zora Hanáčková ◽  
Milada Čiamporová

AbstractUltrastructure was investigated along the files of developing epidermal cells in the root tip of a model plant Medicago sativa, in which all rhizodermal cells are potential hair-forming trichoblasts. Differentiation at subcellular level was observed up to the stage of bulge initiation in the trichoblasts. Root hair initiation indicated by the emergence of bulges from trichoblasts was detected at various distances from the root tip and, it was independent of the trichoblast size.During rhizodermal cell differentiation, starch grains accumulated in the plastids. Nuclei located in the central part of the young, meristematic cells moved towards the inner periclinal wall as the central vacuole enlarged. The bulging region of the trichoblasts located opposite the nucleus and was rich in mitochondria, ER, ribosomes, and Golgi bodies, and contained also vesicles enclosing fibrillar material. This material responded positively to phosphotungstic acid, which was used for detection of cell wall polysaccharides. The cell wall thickness within the bulging domain was significantly lower than in other parts of trichoblasts. We suggest that internalization of cell wall polysaccharides occurs within the bulging area, contributing to local thinning of the cell wall and providing a source of osmotically active compounds for maintaining turgor in the trichoblast. Thus, the internalization process might be necessary for root hair outgrowth.


1996 ◽  
Vol 109 (10) ◽  
pp. 2453-2460 ◽  
Author(s):  
K. Fujimoto ◽  
M. Umeda ◽  
T. Fujimoto

We propose the use of membrane splitting by freeze-fracture for differential phospholipid analysis of protoplasmic and exoplasmic membrane leaflets (halves). Unfixed cells or tissues are quick-frozen, freeze-fractured, and platinum-carbon (Pt/C) shadowed. The Pt/C replicas are then treated with 2.5% sodium dodecyl sulfate (SDS) to solubilize unfractured membranes and to release cytoplasm or contents. While the detergent dissolves unfractured membranes, it would not extract lipids from split membranes, as their apolar domains are stabilized by their Pt/C replicas. After washing, the Pt/C replicas, along with attached protoplasmic and exoplasmic membrane halves, are processed for immunocytochemical labeling of phospholipids with antibody, followed by electron microscopic observation. Here, we present the application of the SDS-digested freeze-fracture replica labeling (SDS-FRL) technique to the transmembrane distribution of a major membrane phospholipid, phosphatidylcholine (PC), in various cell and intracellular membranes. Immunogold labeling revealed that PC is exclusively localized on the exoplasmic membrane halves of the plasma membranes, and the intracellular membranes of various organelles, e.g. nuclei, mitochondria, endoplasmic reticulum, secretory granules, and disc membranes of photoreceptor cells. One exception to this general scheme was the plasma membrane forming the myelin sheath of neurons and the Ca(2+)-treated erythrocyte membranes. In these cell membranes, roughly equal amounts of immunogold particles for PC were seen on each outer and inner membrane half, implying a symmetrical transmembrane distribution of PC. Initial screening suggests that the SDS-FRL technique allows in situ analysis of the transmembrane distribution of membrane lipids, and at the same time opens up the possibility of labeling membranes such as intracellular membranes not normally accessible to cytochemical labels without the distortion potentially associated with membrane isolation procedures.


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