The ultrastructure of Entamoeba sp. (Laredo isolate). Observations on thin sections and freeze-fracture preparations

1979 ◽  
Vol 57 (9) ◽  
pp. 1723-1735 ◽  
Author(s):  
Hampik S. Injeyan ◽  
Erwin Huebner
Keyword(s):  
Particle Density ◽  
Freeze Fracture ◽  
Structural Complexity ◽  
Structural Features ◽  
Nuclear Pore ◽  
Pore Density ◽  
Thin Sections ◽  
Membrane Surfaces ◽  
Intranuclear Microtubules ◽  
First Time

The ultrastructure of Entamoeba sp. (Laredo isolate), seen in thin section and in freeze-fracture preparation, indicates that numerous structural features are common to both this E. histolytica-like amoeba and "regular" strains. In addition, heavy meromyosin (HMM)-binding, actin-like microfilaments and intranuclear microtubules and microfilaments are demonstrated for the first time in Entamoeba. Cytoplasmic microtubules were not detected using two different fixation procedures. Replicas of freeze-fractured membrane surfaces revealed particle size and distribution to be heterogeneous on both fracture faces P and E and suggested a similar structural complexity to that previously described for the HK9 strain of E. histolytica. Particle density was determined in plasma and phagosome membranes, and an enrichment of 4.5-fold and 1.6-fold was estimated for the P and E faces, respectively, of phagosome membranes.Nuclear-pore distribution was heterogenous and pore density was highly variable in log-phase cells probably reflecting different cell-cycle stages. Possible differences in nuclear-pore density and distribution relative to published accounts on "regular" strains of E. histolytica are considered.

scholarly journals Thrombocytopoiesis--analysis by membrane tracer and freeze-fracture studies on fresh human and cultured mouse megakaryocytes.

1984 ◽  
Vol 99 (2) ◽  
pp. 390-402 ◽  
Author(s):  
D Zucker-Franklin ◽  
S Petursson
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Freeze Fracture ◽  
Precursor Cell ◽  
Membrane Domains ◽  
Thin Sections ◽  
Energy Dependent ◽  
First Time ◽  
Later Phase ◽  
Peripheral Cytoplasm

The origin of platelets (Pt) from megakaryocytes (MK) is beyond question, but the mechanism whereby Pts are released from the precursor cell is still debated. A widely-held theory claims that the MK plasma membrane invaginates to form demarcation membranes (DMS), which delineate Pt territories. Accordingly, Pts would be derived mostly from the periphery of the MK, and the MK and Pt plasma membranes would have to be virtually identical. Since, on morphologic grounds, this theory is untenable, several aspects of thrombocytopoiesis were reexamined with the help of membrane tracer and freeze-fracture analyses of freshly-collected human and cultured mouse MK. To our surprise, freeze-cleavage of the MK plasma membrane revealed that the vast majority of intramembranous particles (IMP) remained associated with the protoplasmic leaflet (P face), whereas the partition coefficient of IMPs of the platelet membrane was the reverse. This is the first time that any difference between MK and Pt membranes has been determined. Replicas of freeze-fractured MK that were in the process of thrombocytopoiesis revealed an additional novel phenomenon, i.e., numerous areas of membrane discontinuity that appeared to be related to Pt discharge. When such areas were small, the IMP were lined up along the margin of the crevice. At a later phase, a labyrinth of fenestrations was observed. Thin sections of MK at various stages of differentiation showed that Pt territories were fully demarcated before connections of the DMS with the surface could be found. Therefore, the Pt envelope is probably not derived from invaginations of the MK plasma membrane. When living, MK were incubated with cationic ferritin or peroxidase at 37 degrees C, the tracers entered into the DMS but did not delineate all membranes with which the DMS was in continuity, suggesting the existence of distinctive membrane domains. Interiorization of tracer was not energy-dependent, but arrested at low temperatures. At 4 degrees C the DMS remained empty, unless there was evidence that Pts had been released. In such instances, the tracers outlined infoldings of peripheral cytoplasm that was devoid of organelles. Thus, the majority of Pts seem to originate from the interior of the MK, and the surface membranes of the two cells differ in origin and structure. The observations do not only throw new light on the process of thrombocytopoiesis, but also strengthen the possibility that MKs and Pts may be subject to different stimuli.

scholarly journals Membrane asymmetry and enhanced ultrastructural detail of sarcoplasmic reticulum revealed with use of tannic acid.

1978 ◽  
Vol 79 (3) ◽  
pp. 601-616 ◽  
Author(s):  
A Saito ◽  
C T Wang ◽  
S Fleischer
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Tannic Acid ◽  
Freeze Fracture ◽  
Membrane Vesicles ◽  
Thin Sections ◽  
Membrane Asymmetry ◽  
The Asymmetry ◽  
First Time ◽  
Negative Staining Electron Microscopy

Fixation of purified sarcoplasmic reticulum (SR) membrane vesicles, using glutaraldehyde supplemented with 1% tannic acid, reveals newly visualized ultrastructure in thin sections. The trilaminar appearance of the membrane is highly asymmetric; the outer electron-opaque layer is appreciably wider (70 A) than the inner layer (20 A). The asymmetry is not referable to lack of penetration of the tannic acid since: (a) SR vesicles made permeable with 1 mM EDTA, pH 8.5, show similar asymmetry; (b) treatment of SR with trypsin results in progressive loss in protein content and decrease in the thickness of the outer layer, until in the limit the trilayer has a symmetric appearance; (c) within the same muscle section, the SR membrane appears highly asymmetric whereas the sarcolemma has a more symmetric appearance; (d) reconstituted SR vesicles have a symmetric appearance with equally broad inner and outer layers (approximately 70 A); the symmetric structure is confirmed by freeze-fracture and negative staining electron microscopy. Heavy and light SR vesicles obtained by isopycnic density sedimentation of purified SR have the same asymmetric appearance of the membrane and seem to differ mainly in that the heavy vesicles contain internal contents consisting largely of Ca++-binding protein. The asymmetry of the SR membrane is referable mainly to the unidirectional alignment of the Ca++ pump protein, the major component (90% of the protein) of the membrane. The asymmetry of the SR membrane can be visualized now for the first time in situ in thin sections of muscle.

Stereological Determination of Nuclear Pore Density from Thin Sections of Nuclear Membrane

1974 ◽  
Vol 32 ◽  
pp. 274-275
Author(s):  
Alen V. Loud
Keyword(s):  
Independent Method ◽  
Nuclear Pore ◽  
Pore Density ◽  
Nuclear Pores ◽  
Thin Sections ◽  
Liver Parenchymal ◽  
Normal Rat ◽  
Parenchymal Cells

The number of nuclear pores per square micron of nuclear envelope, NA, has been measured in tangential sections, freeze-etched preparations and isolated fragments of membrane. Estimates of NA for normal rat liver parenchymal cells have varied with the methodology used and range from 14 in freeze-etched nuclei in situ to 36 in negatively stained membranes. An entirely new and independent method for masuring NA, based on the principles of stereology, is presented here.

Membrane Specializations in Developing Nodes of Ranvier

1980 ◽  
Vol 38 ◽  
pp. 626-627
Author(s):  
J.H. Tao-Cheng ◽  
J. Rosenbluth
Keyword(s):  
Schwann Cell ◽  
Freeze Fracture ◽  
Structural Features ◽  
Nerve Fibers ◽  
Nodes Of Ranvier ◽  
Myelinated Nerve ◽  
Thin Sections ◽  
Frog Tadpole ◽  
Cell Layers ◽  
Mature Node

Mature myelinated nerve fibers exhibit distinctive structural features at nodes of Ranvier and the adjacent paranodal regions. In order to obtain information about the interrelationships between these specializations during development, thin sections and freeze-fracture replicas of immature peripheral nerve fibers from grass frog tadpole hind legs were examined during the period of myelinogenesis. Early in myelination axons are enwrapped individually by a few loose Schwann cell layers whose edges overhang each other forming "terminal loops" against the axolemma. Unlike those of the mature node, these loops are widely sepa-rated and irregularly spaced (Fig.l), and similarly the presumptive nodal region between successive developing myelin segments is usually much longer than adult nodes of Ranvier. The presumptive nodal axolemma may exhibit a cytoplasmic "undercoating." However, the density of this coating is highly variable. Usually it is much lower than at adult nodes, and in some cases the undercoating is not distinguishable. The outermost layers of the Schwann cell are usually the first to form axoglial junctional specializations character¬ized by the presence of "transverse bands" and ER cisternae applied to the junctional Schwann cell membrane. In some instances the outermost layer con¬tacts the axon over an extensive area and forms multiple small junctional specializations at widely separated intervals along the length of the axolemma.

Gap junctions in the prothoracic glands of the tobacco hornworm, Manduca sexta

1992 ◽  
Vol 50 (1) ◽  
pp. 706-707
Author(s):  
Ji-da Dai ◽  
M. Joseph Costello ◽  
Lawrence I. Gilbert
Keyword(s):  
Gap Junctions ◽  
Small Molecules ◽  
Manduca Sexta ◽  
Freeze Fracture ◽  
Cell Communication ◽  
Cellular Level ◽  
Thin Sections ◽  
Prothoracic Glands ◽  
Polyhydroxylated Steroids ◽  
Stimulation Of

Insect molting and metamorphosis are elicited by a class of polyhydroxylated steroids, ecdysteroids, that originate in the prothoracic glands (PGs). Prothoracicotropic hormone stimulation of steroidogenesis by the PGs at the cellular level involves both calcium and cAMP. Cell-to-cell communication mediated by gap junctions may play a key role in regulating signal transduction by controlling the transmission of small molecules and ions between adjacent cells. This is the first report of gap junctions in the PGs, the evidence obtained by means of SEM, thin sections and freeze-fracture replicas.

Observations of Frozen-Hydrated Microtubules

1990 ◽  
Vol 48 (1) ◽  
pp. 504-505
Author(s):  
D. Chrétien ◽  
D. Job ◽  
R.H. Wade
Keyword(s):  
Fourier Transforms ◽  
Structural Complexity ◽  
Image Contrast ◽  
Phase Behaviour ◽  
Experimental Conditions ◽  
Thin Sections ◽  
Surface Lattice ◽  
Cilia And Flagella ◽  
Outstanding Question

Microtubules are filamentary structures found in the cytoplasm of eukaryotic cells, where, together with actin and intermediate filaments, they form the components of the cytoskeleton. They have many functions and show various levels of structural complexity as witnessed by the singlet, doublet and triplet structures involved in the architecture of centrioles, basal bodies, cilia and flagella. The accepted microtubule model consists of a 25 nm diameter hollow tube with a wall made up of 13 paraxial protofilaments (pf). Each pf is a string of aligned tubulin dimers. Some results have suggested that the pfs follow a superhelix. To understand how microtubules function in the cell an accurate model of the surface lattice is one of the requirements. For example the 9x2 architecture of the axoneme will depend on the organisation of its component microtubules. We should also note that microtubules with different numbers of pfs have been observed in thin sections of cellular and of in-vitro material. An outstanding question is how does the surface lattice adjust to these different pf numbers?We have been using cryo-electron microscopy of frozen-hydrated samples to study in-vitro assembled microtubules. The experimental conditions are described in detail in this reference. The results obtained in conjunction with thin sections of similar specimens and with axoneme outer doublet fragments have already allowed us to characterise the image contrast of 13, 14 and 15 pf microtubules on the basis of the measured image widths, of the the image contrast symmetry and of the amplitude and phase behaviour along the equator in the computed Fourier transforms. The contrast variations along individual microtubule images can be interpreted in terms of the geometry of the microtubule surface lattice. We can extend these results and make some reasonable predictions about the probable surface lattices in the case of other pf numbers, see Table 1. Figure 1 shows observed images with which these predictions can be compared.

scholarly journals Observations on early fungal infections with relevance for replant disease in fine roots of the rose rootstock Rosa corymbifera 'Laxa'

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
G. Grunewaldt-Stöcker ◽  
C. Popp ◽  
A. Baumann ◽  
S. Fricke ◽  
M. Menssen ◽  
...  
Keyword(s):  
Fine Roots ◽  
Woody Plant ◽  
Fine Root ◽  
Fungal Infections ◽  
Thin Sections ◽  
Replant Disease ◽  
Pot Trials ◽  
Worldwide Phenomenon ◽  
First Time ◽  
The Rose

AbstractReplant disease is a worldwide phenomenon affecting various woody plant genera and species, especially within the Rosaceae. Compared to decades of intensive studies regarding replant disease of apple (ARD), the replant disease of roses (RRD) has hardly been investigated. The etiology of RRD is also still unclear and a remedy desperately needed. In greenhouse pot trials with seedlings of the RRD-sensitive rootstock Rosa corymbifera ‘Laxa’ cultured in replant disease affected soils from two different locations, early RRD symptom development was studied in fine roots. In microscopic analyses we found similarities to ARD symptoms with regards to structural damages, impairment in the root hair status, and necroses and blackening in the cortex tissue. Examinations of both whole mounts and thin sections of fine root segments revealed frequent conspicuous fungal infections in association with the cellular disorders. Particularly striking were fungal intracellular structures with pathogenic characteristics that are described for the first time. Isolated fungi from these tissue areas were identified by means of ITS primers, and many of them were members of the Nectriaceae. In a next step, 35 of these isolates were subjected to a multi-locus sequence analysis and the results revealed that several genera and species were involved in the development of RRD within a single rose plant. Inoculations with selected single isolates (Rugonectria rugulosa and Ilyonectria robusta) in a Perlite assay confirmed their pathogenic relationship to early necrotic host plant reactions, and symptoms were similar to those exhibited in ARD.

Bronze Age Cremation Cemeteries in the East Midlands

1987 ◽  
Vol 53 (1) ◽  
pp. 187-221 ◽  
Author(s):  
Carol S. M. Allen ◽  
Mary Harman ◽  
Hazel Wheeler
Keyword(s):  
Bronze Age ◽  
Direct Relationship ◽  
Small Cluster ◽  
Linear Arrangement ◽  
Thin Sections ◽  
Probable Function ◽  
East Midlands ◽  
Materials Used ◽  
First Time ◽  
Organic Remains

Two Bronze Age cremation cemeteries excavated between 1968 and 1975 are reported and discussed. At Coneygre Farm, Notts., fifty-one cremations were excavated, thirty-one in pots, six in cists, and fourteen uncontained. Cremations were deposited in a roughly linear arrangement and no barrow was found. At Pasture Lodge Farm, Lincs., twenty-seven pots were found, of which twenty-five had associated cremations, and fifteen further sherds could represent burials. Vessels in this cemetery form a small cluster. Pottery from these two cemeteries is broadly similar to Deverel-Rimbury ware and with vessels from other sites in the region is considered to form an East Midlands group of Bronze Age pottery. Vessels of this type from Frieston and Grantham, Lincs., are illustrated for the first time. Examination of thin sections of the pottery from the two cemeteries suggests that most, although not all, of the materials used could have been found locally. Organic remains found in thin sections provide environmental information. The effect of soils on durability of pots and their probable function is discussed. A direct relationship is noticed for the first time between the age of the cremated individual and the capacity of the pot in which the cremation was deposited.

Multi-lamellar organization of fully deuterated lipid extracts of yeast membranes

2014 ◽  
Vol 70 (12) ◽  
pp. 3167-3176 ◽  
Author(s):  
Yuri Gerelli ◽  
Alexis de Ghellinck ◽  
Juliette Jouhet ◽  
Valérie Laux ◽  
Michael Haertlein ◽  
...  
Keyword(s):  
Relative Humidity ◽  
Neutron Diffraction ◽  
Structural Complexity ◽  
Structural Features ◽  
Metabolic Effects ◽  
Growth Media ◽  
Small Momentum Transfer ◽  
Lipid Species ◽  
Two Samples ◽  
Temperature And Humidity

Neutron scattering studies on mimetic biomembranes are currently limited by the low availability of deuterated unsaturated lipid species. In the present work, results from the first neutron diffraction experiments on fully deuterated lipid extracts from the yeastPichia pastorisare presented. The structural features of these fully deuterated lipid stacks are compared with those of their hydrogenous analogues and with other similar synthetic systems. The influence of temperature and humidity on the samples has been investigated by means of small momentum-transfer neutron diffraction. All of the lipid extracts investigated self-assemble into multi-lamellar stacks having different structural periodicities; the stacking distances are affected by temperature and humidity without altering the basic underlying arrangement. At high relative humidity the deuterated and hydrogenous samples are similar in their multi-lamellar arrangement, being characterized by two main periodicities of ∼75 and ∼110 Å reflecting the presence of a large number of polar phospholipid molecules. Larger differences are found at lower relative humidity, where hydrogenous lipids are characterized by a larger single lamellar structure than that observed in the deuterated samples. In both cases the heterogeneity in composition is reflected in a wide structural complexity. The different behaviour upon dehydration can be related to compositional differences in the molecular composition of the two samples, which is attributed to metabolic effects related to the use of perdeuterated growth media.

scholarly journals Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components.

1990 ◽  
Vol 110 (4) ◽  
pp. 883-894 ◽  
Author(s):  
R Reichelt ◽  
A Holzenburg ◽  
E L Buhle ◽  
M Jarnik ◽  
A Engel ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Nuclear Envelope ◽  
Three Dimensional ◽  
Structural Features ◽  
Nuclear Pore ◽  
Xenopus Laevis Oocyte ◽  
Three Dimensional Model ◽  
Transmission Electron ◽  
Freeze Dried

Nuclear pore complexes (NPCs) prepared from Xenopus laevis oocyte nuclear envelopes were studied in "intact" form (i.e., unexposed to detergent) and after detergent treatment by a combination of conventional transmission electron microscopy (CTEM) and quantitative scanning transmission electron microscopy (STEM). In correlation-averaged CTEM pictures of negatively stained intact NPCs and of distinct NPC components (i.e., "rings," "spoke" complexes, and "plug-spoke" complexes), several fine structural features arranged with octagonal symmetry about a central axis could reproducibly be identified. STEM micrographs of unstained/freeze-dried intact NPCs as well as of their components yielded comparable but less distinct features. Mass determination by STEM revealed the following molecular masses: intact NPC with plug, 124 +/- 11 MD; intact NPC without plug, 112 +/- 11 MD; heavy ring, 32 +/- 5 MD; light ring, 21 +/- 4 MD; plug-spoke complex, 66 +/- 8 MD; and spoke complex, 52 +/- 3 MD. Based on these combined CTEM and STEM data, a three-dimensional model of the NPC exhibiting eightfold centrosymmetry about an axis perpendicular to the plane of the nuclear envelope but asymmetric along this axis is proposed. This structural polarity of the NPC across the nuclear envelope is in accord with its well-documented functional polarity facilitating mediated nucleocytoplasmic exchange of molecules and particles.

Sign in / Sign up

Export Citation Format

Share Document