?Cholinergic? postsynaptic membranes of bullfrog sympathetic ganglia: Electron microscopy of thin sections and freeze-fracture replicas

1986 ◽  
Vol 214 (1) ◽  
pp. 82-88 ◽  
Author(s):  
Hiroshi Watanabe ◽  
Hiroshi Washioka ◽  
Akira Tonosaki
Keyword(s):  
Electron Microscopy ◽  
Freeze Fracture ◽  
Sympathetic Ganglia ◽  
Thin Sections

scholarly journals "Reticular" and "Areticular" Nissl Bodies in Sympathetic Neurons of a Lizard

1959 ◽  
Vol 6 (1) ◽  
pp. 77-84 ◽  
Author(s):  
Stuart W. Smith
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Sympathetic Neurons ◽  
Sympathetic Ganglia ◽  
Thin Sections ◽  
Phrynosoma Cornutum ◽  
Reticular Structure ◽  
Before And After ◽  
Partial Dispersion ◽  
Nissl Substance

Sympathetic ganglia of the horned lizard, Phrynosoma cornutum, were fixed in OsO4 and imbedded in methacrylate. Thin sections were cut for electron microscopy. Some adjacent thick sections were cut for light microscopy and were stained in acidified, dilute thionine both before and after digestion by RNase. In the light microscope two types of Nissl bodies are found, both removable by RNase: (1) a deep, diffuse, indistinctly bounded, metachromatic variety, and (2) a superficial, dense, sharply delimited, orthochromatic sort. Electron microscopically, the former ("reticular" Nissl bodies) corresponds to the granulated endoplasmic reticular structure of Nissl material previously described by others, whereas the latter ("areticular" Nissl bodies) comprises compact masses of particles of varying internal density and devoid of elements of endoplasmic reticulum. The constituent particles of the areticular Nissl material are 4 to 8 x the diameter of single ribonucleoprotein granules of the reticular Nissl substance and seem, near zones of junction with the reticular type, to arise by clustering of such granules with subsequent partial dispersion of the substance of the granules into an added, less dense material. It is suggested that the observed orthochromasia of the areticular Nissl substance is due to accumulation of a large amount of protein bound to RNA and, further, that these Nissl bodies may represent storage depots of RNA and protein.

Differentiation in Trypanosoma brucei: host-parasite cell junctions and their persistence during acquisition of the variable antigen coat

1985 ◽  
Vol 74 (1) ◽  
pp. 1-19 ◽  
Author(s):  
L. Tetley ◽  
K. Vickerman
Keyword(s):  
Electron Microscopy ◽  
Trypanosoma Brucei ◽  
Morphological Changes ◽  
Freeze Fracture ◽  
Tsetse Fly ◽  
Cell Junctions ◽  
Surface Coat ◽  
Thin Sections ◽  
Variable Antigen ◽  
Host Parasite

Acquisition of the variable antigen-containing surface coat of Trypanosoma brucei occurs at the metacyclic stage in the salivary glands of the tsetse fly vector. The differentiation of the metacyclic trypanosome in the gland has been studied by scanning electron microscopy and by transmission electron microscopy of thin sections and freeze-fracture replicas. The uncoated epimastigote trypanosomes (with a prenuclear kinetoplast) divide while attached to the salivary gland epithelium brush border by elaborate branched flagellar outgrowths, which ramify between the host cell microvilli and form punctate hemidesmosome-like attachment plaques where they are indented by the microvilli. These outgrowths become reduced as the epimastigotes transform to uncoated trypomastigotes (with postnuclear kinetoplast), which remain attached and capable of binary fission. The flagellar outgrowths disappear but the attachment plaques persist as the uncoated trypomastigotes (premetacyclics) stop dividing and acquire the surface coat to become ‘nascent metacyclics’. Coat acquisition therefore occurs in the attached trypanosome and not, as previously believed, after detachment. Coating is accompanied by morphological changes in the glycosomes and mitochondrion of the parasite. Freeze-fracture replicas of the host-parasite junctional complexes show membrane particle aggregates on the host membrane but not on the parasite membrane. It is suggested that disruption of the complex occurs when maximum packing of the glycoprotein molecules has been achieved in the trypanosome surface coat, releasing the metacyclic trypanosome into the lumen of the gland.

scholarly journals Formation of multilamellar vesicles by addition of tannic acid to phosphatidylcholine-containing small unilamellar vesicles.

1989 ◽  
Vol 37 (11) ◽  
pp. 1635-1643 ◽  
Author(s):  
A H Schrijvers ◽  
P M Frederik ◽  
M C Stuart ◽  
K N Burger ◽  
V V Heijnen ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Tannic Acid ◽  
Freeze Fracture ◽  
Tissue Preparation ◽  
Unilamellar Vesicles ◽  
Thin Sections ◽  
Multilamellar Vesicles ◽  
Morphological Studies ◽  
Rat Hearts ◽  
Small Unilamellar Vesicles

Tannic acid induces aggregation and formation of multilamellar vesicles when added to preparations of small unilamellar vesicles, specifically those containing phosphatidylcholine. Aggregation and clustering of vesicles was demonstrated by cryo-electron microscopy of thin films and by freeze-fracture technique. Turbidity measurements revealed an approximately one-to-one molar ratio between tannic acid and phosphatidylcholine necessary for a fast and massive aggregation of the small unilamellar vesicles. When tannic acid-induced aggregates were dehydrated and embedded for conventional thin-section electron microscopy, multilamellar vesicles were retrieved in thin sections. It is concluded from morphological studies, as well as previous tracer studies, that tannic acid, at least to a great extent, prevents the extraction of phosphatidylcholine. Multilamellar vesicles were also observed in tannic acid-treated vesicles prepared from total lipid extracts from either rabbit or rat hearts. Substantially more multilamellar vesicles were retrieved in the rabbit vesicle preparation. This difference can probably be explained by the difference in the proportion of the plasmalogen phosphatidylcholine, and possibly the content of sphingomyelin, in lipid extracts of rabbit and rat hearts. It is concluded that the dual effect (reduced extraction and aggregation) of tannic acid on phosphatidylcholines should be taken into consideration when tannic acid is used in tissue preparation.

scholarly journals Evidence of a Tubular System for Transendothelial Transport in Arterial Capillaries of the Rete Mirabile

1997 ◽  
Vol 45 (10) ◽  
pp. 1365-1378 ◽  
Author(s):  
Moöse Bendayan ◽  
Eugenio A. Rasio
Keyword(s):  
Electron Microscopy ◽  
Serum Proteins ◽  
Protein A ◽  
Freeze Fracture ◽  
Thin Sections ◽  
Functional Studies ◽  
Transendothelial Transport ◽  
Rete Mirabile ◽  
Osmium Impregnation ◽  
Membrane Bound

The arterial endothelial cells of the rete capillaries of the eel were examined by transmission electron microscopy on thin sections, on freeze-fracture replicas, by scanning electron microscopy, after cytochemical osmium impregnation and perfusion with peroxidase. The study revealed the existence of membrane-bound tubules and vesicles that open at both the luminal and abluminal poles of the cell and at the level of the intercellular space. The tubules are straight or present successive dilations and constrictions. They branch in various directions and intrude deeply into the cell cytoplasm, forming a complex tubular network within the cell. Immunocytochemical techniques were applied on immersion-fixed tissues and on perfusion of the capillaries with albumin and insulin. These demonstrated that the tubular–vesicular system is involved in the transport of circulating proteins. Furthermore, protein A–gold immunocytochemistry has revealed the association of actin with the membranes of this system. On the basis of these results, we suggest that the transendothelial transport of serum proteins takes place by a transcytotic process through a membrane-bound tubular–vesicular system and is equivalent to the large pore system presumed from functional studies.

The extrarhabdomeral cytoskeleton in photoreceptors of Diptera. II. Plasmalemmal undercoats

1984 ◽  
Vol 220 (1220) ◽  
pp. 353-359 ◽  
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Genetic Analysis ◽  
Freeze Fracture ◽  
Integral Membrane Proteins ◽  
Basal Region ◽  
Thin Sections ◽  
En Face ◽  
Thiol Proteases ◽  
Planar Membranes

The plasmalemmal undercoats of those regions of the photoreceptors of the blowfly Lucilia that flank the central extracellular space of each ommatidium are described from en face and transverse thin sections. Labile structures were stabilized before fixation for electron microscopy by using an inhibitor of thiol proteases, Ep-475, as described in the previous paper (Blest et al., Proc. R. Soc. Lond . B 220, 339-352, 1984). Membranes of R 1-6 are underlain by a closely associated, randomly organized filamentous meshwork. That of the basal region of R 7 is highly organized, and consists of very long, about 8 nm filaments running parallel to each other and to the longitudinal ommatidial axis; these ‘backbone’ filaments are tightly adherent to the plasma membrane, and are spaced some 190-200 nm apart. They are linked by abundant transverse filaments that form a reticulum between them. The degree of ordering of the reticulum in life is not clear, but some well-preserved profiles suggest that it may be high. Replicas obtained by the freeze-fracture technique show that extrarhabdomeral membranes have dense populations of intramembrane particles, just as they do in Drosophila where a genetic analysis has shown them to consist largely of rhodopsin. It is proposed as a working hypothesis that these planar membranes can be regarded as flat equivalents of the microvillar membranes, that some fraction of the integral membrane proteins may be immobilized by bonding to the plasmalemmal undercoat, and that the latter may help to constrain both the translational and rotational movements of rhodopsin molecules.

scholarly journals Ultrastructure of Na,K-transport vesicles reconstituted with purified renal Na,K-ATPase.

1980 ◽  
Vol 86 (3) ◽  
pp. 746-754 ◽  
Author(s):  
E Skriver ◽  
A B Maunsbach ◽  
P L Jørgensen
Keyword(s):  
Electron Microscopy ◽  
Freeze Fracture ◽  
Cation Transport ◽  
Coupled Transport ◽  
Vesicle Membrane ◽  
Thin Sections ◽  
Intramembrane Particles ◽  
Freeze Fracture Electron Microscopy ◽  
Fracture Electron ◽  
K Transport

To study the size and structure of the Na,K-pump molecule, the ultrastructure of phospholipid vesicles was examined after incorporation of purified Na,K-ATPase which catalyzes active coupled transport of Na+ and K+ in a ratio close to 3Na/2K. The vesicles were analyzed by thin sectioning and freeze-fracture electron microscopy after reconstitution with different ratios of Na,K-ATPase protein to lipid, and the ultrastructural observations were correlated to the cation transport capacity. The purified Na,K-ATPase reconstituted with phospholipids to form a very uniform population of vesicles. Thin sections of preparations fixed with glutaraldehyde and osmium tetroxide showed vesicles limited by a single membrane which in samples stained with tannic acid appeared triple-layered with a thickness of 70 A. Also, freeze-fracture electron microscopy demonstrated uniform vesicles with diameters in the range of 700-1,100 A and an average value close to 900 A. The vesicle diameter was independent of the amount of protein used for reconstitution. Intramembrane particles appeared only in the vesicle membrane after introduction of Na,K-ATPase and the frequency of intramembrane particles was proportional to the amount of Na,K-ATPase protein used in the reconstitution. The particles were evenly distributed on the inner and the outer leaflet of the vesicle membrane. The diameter of the particles was 90 A and similar to our previous values for the diameter of intramembrane particles in the purified Na,K-ATPase. The capacity for active cation transport in the reconstituted vesicles was proportional to the frequency of intramembrane particles over a range of 0.2-16 particles per vesicle. The data therefore show that active coupled Na,K transport can be carried out by units of Na,K-ATPase which appear as single intramembrane particles with diameters close fo 90 A in the freeze-fracture micrographs.

scholarly journals Observations on the mitochondrial reticulum in the yeast Candida utilis as revealed by freeze-fracture electron microscopy

1980 ◽  
Vol 46 (1) ◽  
pp. 289-297
Author(s):  
E. Keyhani
Keyword(s):  
Electron Microscopy ◽  
Cross Sections ◽  
Candida Utilis ◽  
Freeze Fracture ◽  
Yeast Cells ◽  
Logarithmic Phase ◽  
Thin Sections ◽  
Freeze Fracture Electron Microscopy ◽  
Fracture Electron ◽  
Mitochondrial Reticulum

Freeze-fracture of Candida utilis yeast cells grown to early logarithmic phase (5 h) and stationary phase (24 h) revealed a branched mitochondrial reticulum both in longitudinal and transverse cross-fracture studies. In contrast, the longitudinal and transverse sections of permanganate-fixed cells showed that the small ovoid or spherical mitochondria were located at the periphery of the cell, without formation of a reticulum. Data indicate that the separate mitochondrial profiles seen in thin sections of permanganate-fixed yeast cells are not separate round or ovoid mitochondria, but rather are cross-sections of the mitochondrial reticulum.

Ultrastructure of the dormant and germinating sporangiospore of Phascolomyces articulosus (Mucorales)

1978 ◽  
Vol 56 (7) ◽  
pp. 747-753 ◽  
Author(s):  
P. Jeffries ◽  
T. W. K. Young
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Freeze Fracture ◽  
Spore Wall ◽  
Wall Layer ◽  
Thin Sections ◽  
Transmission Electron ◽  
Sporangial Wall ◽  
Scanning Electron

Using results obtained with light and scanning electron microscopy of critical-point-dried material and transmission electron microscopy of carbon replicas and freeze-fracture and ultra-thin sections, the structure and germination of the sporangiospore of Phascolomyces articulosus Boedijn is described. The sporangial wall is trilaminate and the ornamented spore wall is two layered. During germination, a new wall layer develops between the plasmalemma and the original spore wall. Sporangial structure is related to that of other members of the Thamnidiaceae and the use of germinating spores of P. articulosus for infection studies of the mycoparasite Piptocephalis unispora is indicated.

Intercellular junctions in the larval epidermis of the colonial ascidian Distaplia occidentalis: a thin-section and freeze-fracture examination

1986 ◽  
Vol 64 (1) ◽  
pp. 112-117 ◽  
Author(s):  
Michael J. Cavey ◽  
Richard L. Wood
Keyword(s):  
Electron Microscopy ◽  
Freeze Fracture ◽  
Intercellular Junctions ◽  
Epidermal Cells ◽  
Cellular Membranes ◽  
Colonial Ascidian ◽  
Thin Sections ◽  
Intramembranous Particles ◽  
Transmission Electron ◽  
Columnar Cells

The larval epidermis of the colonial ascidian Distaplia occidentalis is a unilayered epithelium consisting of squamous and cuboidal or low columnar cells. The epidermal cells are laterally folded and interdigitated or overlapped. The occluding (tight) junctions and the close (gap) junctions that join the epidermal cells have been examined by transmission electron microscopy. In thin sections, the occluding junction is represented by focal fusions of the apposed plasmalemmata. Freeze-fracture replicas of the occluding junction show linear, anastomosing arrays of intramembranous particles on the protoplasmic faces of the cellular membranes. In thin sections of the close junction, the apposed plasmalemmata are mutually parallel and separated by a narrow intercellular cleft. Freeze-fracture replicas of the close junction reveal macular aggregations of intramembranous particles on the protoplasmic faces of the cellular membranes.

Structure of the Stigma and Style of the Avocado

1978 ◽  
Vol 26 (5) ◽  
pp. 663 ◽  
Author(s):  
M Sedgley ◽  
MS Buttrose
Keyword(s):  
Electron Microscopy ◽  
Smooth Endoplasmic Reticulum ◽  
Freeze Fracture ◽  
Persea Americana ◽  
Transmitting Tissue ◽  
Intercellular Substance ◽  
Thin Sections ◽  
Transmission Electron ◽  
Tissue Cells ◽  
Stigma Secretion

The structure of the stigma and style of the avocado (Persea americana Mill.) was investigated by light microscopy, scanning electron microscopy and transmission electron microscopy of thin sections and freeze-fracture replicas. The stigmalstyle was asymmetrical and a groove, lined with transmitting tissue, extended the whole length of the structure. Stigma papillae fringed this groove for about a third of its length. There was no clear distinction between stigma papillae and stylar transmitting tissue cells but there was a gradation of structure down the axis. The papilla cells were long with large and small vacuoles; the transmitting tissue cells had small vacuoles only. The stigma secretion and intercellular substance of the transmitting tissue contained carbohydrate and lipid. Clusters of plastids with little internal structure and electron-dense stroma were abundant in the cells of the stigma and transmitting tissue along with extensive smooth endoplasmic reticulum. Both single vesicles and multivesicular bodies were observed fusing with the plasmalemma which was abnormally rough in freeze-fracture profiles. It is suggested that the cells of the stigma and transmitting tissue have a largely secretory function and may be approaching or have reached senescence when the flower opens.

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