Radioautographic visualization of β-glucans formed by pea membranes from UDP-glucose

1983 ◽  
Vol 61 (4) ◽  
pp. 1266-1275 ◽  
Author(s):  
Susette C. Mueller ◽  
Gordon A. Maclachlan

Radioautographic experiments were carried out using pea stem slices to determine the site of glucose incorporation from UDP-glucose. Cut or damaged pea stem cells were the only cells to incorporate [3H]glucose from UDP-[3H]glucose. The product formed at 20 μM UDP-glucose was observed in electron microscope thin sections in patches on the plasma membrane and the cell wall. The product formed at 5 mM UDP-glucose occurred in fibrillar bundles that stretched between the plasma membrane and the cell wall. This periplasmic material fluoresced when stained with aniline blue. Experiments in which slices were subjected to sequential incubations in radioactive 5 mM UDP-glucose followed by unlabelled 5 mM UDP-glucose, or incubations in the reverse order, indicated that incorporation of [3H]glucose into products insoluble in chloroform:methanol:water or hot water occurs at the plasma membrane, and radioactivity is displaced from the membrane by subsequent incubations. A similar experiment, in which slices were first incubated in radioactive 20 μM UDP-glucose followed by unlabelled 5 mM UDP-glucose, indicated that the synthesis of fibrillar material from 5 mM UDP-glucose displaces the labelled product that had been formed from 20 μM UDP-glucose. It is concluded that only cut or damaged pea stem cells utilize UDP-glucose and the plasma membrane enzymes that incorporate [3H]glucose from 20 μM or 5 mM UDP-[3H]glucose are in close physical proximity.

1964 ◽  
Vol 20 (2) ◽  
pp. 217-233 ◽  
Author(s):  
G. W. Claus ◽  
L. E. Roth

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.


1963 ◽  
Vol 18 (2) ◽  
pp. 429-440 ◽  
Author(s):  
Ryan W. Drum

The cytoplasmic fine structure of the motile, pennate diatom, Nitzschia palea was studied in thin sections viewed in the electron microscope. The cells were fixed in OsO4, embedded in methacrylate, and immersed in 10 per cent hydrofluoric acid (HF) for 36 to 40 hours to remove the siliceous cell wall prior to sectioning. The HF treatment did not cause any obvious cytoplasmic damage. The dictyosome complex is perinuclear, and located only in the central cytoplasm. Mitochondria are sparse in the central cytoplasm, but abundant in the peripheral cytoplasm, and fill many of the transvacuolar cytoplasmic strands. Characteristic, amorphous oil bodies fill certain cytoplasmic strands and probably are not leucosin. The pyrenoid appears to be membrane limited, and oil droplets are found adjacent to the pyrenoid. The pyrenoid of another diatom, Cymbella affinis, is also membrane-limited. The membrane limiting the pyrenoid may be a composite of the terminal portions of chloroplast discs, facilitating rapid movement of photosynthate into the pyrenoid matrix, where the characteristic oil droplets may be formed. Carinal fibrils are found singly in each carinal pore, and may be involved in the locomotion of Nitzschia palea.


1966 ◽  
Vol 12 (6) ◽  
pp. 1125-1136 ◽  
Author(s):  
Alice Reyn ◽  
A. Birch-Andersen ◽  
S. P. Lapage

The line structure of Haemophilus vaginalis (Gardner and Dukes 1955) was compared with that of four, possibly related species (Butyribacterium rettgeri, Corynebacterium diphtheriae var. mitis, Lactobacillus acidophilus, Haemophilus influenzae) and an unrelated species, Neisseria haemolysans, which had shown a similar degree of Gram-variability as that of H. vaginalis. Although H. vaginalis was first described as a Gram-negative rod, its fine structure, particularly that of cell wall and septa, was more like that of Gram-positive organisms. Also N. haemolysans had a fine structure close to that of Gram-positive organisms, and its typical Gram-positive cell wall varied in. thickness from one cell to another.The study did not solve the problem of the classification of the so-called H. vaginalis, but the appearance of the few strains studied in the electron microscope suggests that it: should be included in Corynebacterium or Butyribacterium rather than in Lactobacillus.


1972 ◽  
Vol 18 (7) ◽  
pp. 997-1002 ◽  
Author(s):  
I. L. Stevenson ◽  
S. A. W. E. Becker

Methods have been developed for the rapid, reproducible induction of high-density populations of F. oxysporum chlamydospores. On transferring washed pregerminated conidia to a simple two-salts medium, chlamydospore morphogenesis was evident by 12 h and masses of mature spores could be harvested at the end of 4 days. Electron-microscope studies of thin sections of mature chlamydospores reveal a thick triple-layered cell wall. The cytoplasm contains, in addition to large lipid deposits, a nucleus, mitochondria, and endoplasmic reticulum all typical of fungal cells. Chlamydospores of F. oxysporum exhibit two distinct types of cell surface in thin section. The outer wall layer of two of the isolates studied was smooth-surfaced while the outer layer of the two other isolates was distinctly fibrillose. Some evidence is presented suggesting that the fibrillose material arises through the partial breakdown of the original hyphal wall.


1962 ◽  
Vol 13 (1) ◽  
pp. 109-115 ◽  
Author(s):  
Robert C. Buck ◽  
James M. Tisdale

The development of the mid-body has been studied in mitotic erythroblasts of the rat bone marrow by means of thin sections examined with the electron microscope. A differentiated region on the continuous spindle fibers, consisting of a localized increase in density, is observed at the equatorial plane. The mid-body seems to develop by the aggregation of such denser lengths of spindle fiber. Its appearance precedes that of the cleavage furrow. A plate-like arrangement of fibrillary material lies transversely across the telophase intercellular bridge. Later, this material becomes amorphous and assumes the form of a dense ring closely applied to a ridge in the plasma membrane encircling the middle of the bridge. Although the mid-body forms in association with the spindle fibers, it is a structurally distinct part, and the changes which it undergoes are not shared by the rest of the bundle of continuous fibers.


1968 ◽  
Vol 14 (10) ◽  
pp. 1049-1052 ◽  
Author(s):  
N. E. Dweltz ◽  
J. Ross Colvin

The structure of the diatom, Thalassiosira fluviatilis, was investigated by a study with the electron microscope of thin sections of cells fixed by OsO4, KMnO4, and glutaraldehyde. The weakly siliceous cell wall is composed of a thin sheet perforated by holes about 250 Å in diameter and arranged in a roughly orthogonal manner. At the ends of the barrel-shaped cell are marginal and central pores composed of an outer cylinder (0.25 μ in diameter and 1 μ long) arranged coaxially with an inner, smaller, shorter cylinder. The thin, perforated sheet (valve) which connects these pores is reinforced from place to place by radial or transverse ribs.Mitochondria, nuclei, chloroplasts, and pyrenoids have the same general structure and appearance as those in other diatoms. Small vacuoles are numerous but it is not known whether they are filled with gas or solution. The cells often show aggregates of an amorphous material which does not stain like lipid or protein but which may be the residue of a gelatinous, reserve carbohydrate (chrysolaminarin).


1967 ◽  
Vol 35 (1) ◽  
pp. 37-51 ◽  
Author(s):  
Jack L. Pate ◽  
Erling J. Ordal

An electron microscope study of the myxobacterium Chondrococcus columnaris has revealed the following structures in the peripheral layers of the cells: (1) a plasma membrane, (2) a single dense layer (probably the mucopeptide component of the cell wall), (3) peripheral fibrils, (4) an outer membrane, and (5) a material coating the surfaces of the cells which could be stained with the dye ruthenium red.The ruthenium red-positive material is probably an acid mucopolysaccharide and may be involved in the adhesive properties of the cells. The outer membrane and plasma membrane both have the appearance of unit membranes: an electron-translucent layer sandwiched between two electron-opaque layers. The peripheral fibrils span the gap between the outer membrane and the mucopeptide layer, a distance of about 100 A, and run parallel to each other along the length of the cell. The fibrils appear to be continuous across the ends of the cells. The location of these fibrillar structures suggests that they may play a role in the gliding motility of these bacteria.


1959 ◽  
Vol 6 (3) ◽  
pp. 383-392 ◽  
Author(s):  
H. E. Karrer

The musculature of small lung veins, of the thoracic portion of the inferior vena cava, and of other thoracic veins of the mouse have been studied in the electron microscope. Tissues were fixed in 1 per cent osmium tetroxide buffered with veronal, to which either sodium chloride or sucrose had been added. Methacrylate or araldite served as embedding matrices. Phosphotungstic acid or uranyl acetate was used to stain some of the preparations. Thin sections were examined in a Siemens and Halske Elmiskop Ib electron microscope. The entire musculature of the veins examined was of the striated type. It represents a variety of cardiac muscle, characterized by centrally located nuclei, typical mitochondria, and narrow I bands. Many I bands cannot be recognized at all. H and M bands are likewise indistinct. There is a double array of primary and secondary myofilaments. Mitochondria are large and numerous and contain many cristae. The endoplasmic reticulum consists of longitudinal tubules which run through the whole sarcomeres and bypass Z bands, and of transverse tubules which accompany Z bands. Some "triads," located at Z levels, consist of flattened vacuoles flanked by such transverse tubules. Small vesicles located at Z bands, close to the nucleus, and beneath the plasma membrane may represent still other portions of the reticulum.


1977 ◽  
Vol 23 (4) ◽  
pp. 378-388 ◽  
Author(s):  
J. C. Tu ◽  
S. K. Malhotra

Effect of cyclic AMP (cAMP) on Phycomyces blakesleeanus was studied by growing sporangiospores on glucose–asparagine agar or liquid medium containing three different levels of cAMP (10, 20 and 40 μM) in addition to the control (no cAMP added). The response of Phycomyces to the exogenous cAMP concentration in the medium is as follows: (1) the time required for germ tube emergence is reduced; (2) the diameter of the mycelium is increased (sometimes more than 10 times) and frequency of branching is also increased; (3) the cell wall of the mycelium is thickened (in some cases more than 5 times); (4) the glycogen in the cytoplasm is decreased as visualized in thin sections and also demonstrated in biochemical quantitation; and (5) the distribution of intercalated membranous particles (Imp) on plasma membrane is altered and this can be easily detected in freeze-fractured replica. Such a change in Imp is seen in the formation of small clusters of aggregated particles on the plasmic half (PF) and craters on the complementary exoplasmic half(EF)of the plasma membrane. Although the mechanism of cAMP action requires further exploration, it is possible that the addition of cAMP to the culture medium leads to degradation of glycogen and enhancement of chitin synthesis since the cell wall is largely composed of chitin. The alteration in Imp may be related to a change in the activity of chitin synthetase which is a plasma membrane-bound enzyme.


1978 ◽  
Vol 56 (21) ◽  
pp. 2666-2693 ◽  
Author(s):  
G. B. Ouellette

Seven years (1970–1977) of comparative light and electron microscope studies show that extensive cell wall disruption and breakdown occur consistently in elm xylem tissues infected by Ceratocystis ulmi (Buism.) C. Moreau. These alterations, noticeable even in incipient infection, can be related to the severity of wilting development and occur in association with the presence of an unbound osmiophilic material containing fine fibrillar material, dense particles of approximately 15 nm, and multilamellate structures. Masses of unbound osmiophilic material in host walls and walled fungal cells with which it is sometimes continuous are highly and exclusively labeled following injection of [6-3H]thymidine.The presence of osmiophilic material in host walls and the interrelation between the two was further established by examining stereoscopic pairs of prints taken at various angles with a goniometer. This type of cell wall breakdown seems difficult to relate to other known types of wood rots. Further discussion on the possible nature and origin of the osmiophilic material is presented.


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