Lanthanum and freeze fracture studies on the retinular cell junction in the compound eye of the housefly

1981 ◽  
Vol 214 (3) ◽  
Author(s):  
Che Chi ◽  
StanleyD. Carlson
Keyword(s):  
Compound Eye ◽  
Freeze Fracture ◽  
Cell Junction ◽  
Retinular Cell

The extrarhabdomeral cytoskeleton in photoreceptors of Diptera. I. Labile components in the cytoplasm

1984 ◽  
Vol 220 (1220) ◽  
pp. 339-352 ◽  
Keyword(s):  
Electron Microscopy ◽  
Actin Filaments ◽  
Compound Eye ◽  
Freeze Fracture ◽  
Dynamic Processes ◽  
Ionophore A23187 ◽  
The Status ◽  
Thiol Proteases ◽  
Cytoskeletal Elements ◽  
Nm Range

Labile cytoskeletal structures in the cytoplasm of photoreceptors of the blowfly Lucilia and of Drosophila were stabilized before primary fixation for electron microscopy by retinal infiltration with two inhibitors of thiol proteases, Ep-459 or Ep-475. Alternatively, pretreatments employed EGTA in combination with the Ca 2+ ionophore A23187. The following cytoskeletal structures were revealed, (i) Radial, robust filaments run roughly parallel to the axes of the rhabdomeral microvilli and may be continuous with the axial microvillar filaments. They have diameters of 8 nm upwards, and are occasionally seen to be in association with radial microtubules and with pigment granules. (ii) Slender radial filaments with diameters in the 6-8 nm range extend for shorter distances from the bases of microvilli, and are also associated with endocytotic structures. (iii) The receptor cytoplasm is densely occupied by an ill-defined, filamentous network. (iv) Bundles of slender filaments run longitudinally on each side of rhabdoms of R 1-6 in Lucilia , close to the plasma membrane. Dimensions cited for all categories of filament must be treated with caution because of problems of resolution. Photoreceptors do not bind the fluorescent F-actin probe NBD-phallacidin either without or after treatment with thiol protease inhibitors, and slender filaments are of greater diameter than the 4-5 nm obtained for identified actin filaments in the basement membrane of the compound eye of Lucilia . Infiltration of retinae with Ep-459 or Ep-475 neither prejudices phototransduction, nor impairs the radial migrations of granules of screening pigment in response to light or dark adaption. The status of these cytoskeletal elements is discussed in terms of the dynamic processes of the photoreceptors, and of various labile filaments described from recent studies of vertebrate material using the deep-etch freeze-fracture technique.

Ultrastructural Investigations of the Contractile Filaments of Amoebae

1974 ◽  
Vol 32 ◽  
pp. 188-189
Author(s):  
P.L. Moore
Keyword(s):  
Cytoplasmic Streaming ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Amoeboid Movement ◽  
Different Types ◽  
Chemical Conditions ◽  
Complete Interpretation

Previous freeze fracture results on the intact giant, amoeba Chaos carolinensis indicated the presence of a fibrillar arrangement of filaments within the cytoplasm. A complete interpretation of the three dimensional ultrastructure of these structures, and their possible role in amoeboid movement was not possible, since comparable results could not be obtained with conventional fixation of intact amoebae. Progress in interpreting the freeze fracture images of amoebae required a more thorough understanding of the different types of filaments present in amoebae, and of the ways in which they could be organized while remaining functional.The recent development of a calcium sensitive, demembranated, amoeboid model of Chaos carolinensis has made it possible to achieve a better understanding of such functional arrangements of amoeboid filaments. In these models the motility of demembranated cytoplasm can be controlled in vitro, and the chemical conditions necessary for contractility, and cytoplasmic streaming can be investigated. It is clear from these studies that “fibrils” exist in amoeboid models, and that they are capable of contracting along their length under conditions similar to those which cause contraction in vertebrate muscles.

Photoreceptor disk membrane substructures by means of the complementary freeze-fracture-replica methods

1978 ◽  
Vol 36 (2) ◽  
pp. 156-157
Author(s):  
A. Tonosaki ◽  
M. Yamasaki ◽  
H. Washioka ◽  
J. Mizoguchi
Keyword(s):  
Cell Membranes ◽  
Freeze Fracture ◽  
Purple Membrane ◽  
Remarkable Case ◽  
Single Face ◽  
Disk Membrane ◽  
Associated Proteins ◽  
Photoreceptor Membranes

A vertebrate disk membrane is composed of 40 % lipids and 60 % proteins. Its fracture faces have been classed into the plasmic (PF) and exoplasmic faces (EF), complementary with each other, like those of most other types of cell membranes. The hypothesis assuming the PF particles as representing membrane-associated proteins has been challenged by serious questions if they in fact emerge from the crystalline formation or decoration effects during freezing and shadowing processes. This problem seems to be yet unanswered, despite the remarkable case of the purple membrane of Halobacterium, partly because most observations have been made on the replicas from a single face of specimen, and partly because, in the case of photoreceptor membranes, the conformation of a rhodopsin and its relatives remains yet uncertain. The former defect seems to be partially fulfilled with complementary replica methods.

Decoration of specific sites on freeze-fractured membranes

1978 ◽  
Vol 36 (2) ◽  
pp. 140-141
Author(s):  
H. Gross ◽  
H. Moor
Keyword(s):  
Pure Water ◽  
Freeze Fracture ◽  
Chemical Properties ◽  
Surface Forces ◽  
Sulfate Pentahydrate ◽  
Copper Sulfate Pentahydrate ◽  
Physico Chemical ◽  
Water Of Hydration ◽  
Small Container ◽  
Binding Forces

Fracturing under ultrahigh vacuum (UHV, p ≤ 10-9 Torr) produces membrane fracture faces devoid of contamination. Such clean surfaces are a prerequisite foe studies of interactions between condensing molecules is possible and surface forces are unequally distributed, the condensate will accumulate at places with high binding forces; crystallites will arise which may be useful a probes for surface sites with specific physico-chemical properties. Specific “decoration” with crystallites can be achieved nby exposing membrane fracture faces to water vopour. A device was developed which enables the production of pure water vapour and the controlled variation of its partial pressure in an UHV freeze-fracture apparatus (Fig.1a). Under vaccum (≤ 10-3 Torr), small container filled with copper-sulfate-pentahydrate is heated with a heating coil, with the temperature controlled by means of a thermocouple. The water of hydration thereby released enters a storage vessel.

Freeze-fracture, freeze-etch complementary pairs of virus-infected cells reveal value of etching even when relatively high concentrations of cryoprotectants are used

1978 ◽  
Vol 36 (2) ◽  
pp. 136-137
Author(s):  
Russell L. Steere ◽  
Eric F. Erbe
Keyword(s):  
Plant Tissue ◽  
Phosphate Buffer ◽  
Infected Plant ◽  
Freeze Fracture ◽  
Distilled Water ◽  
Virus Infected Plant ◽  
Infected Cells ◽  
High Concentrations

It has been assumed by many involved in freeze-etch or freeze-fracture studies that it would be useless to etch specimens which were cryoprotected by more than 15% glycerol. We presumed that the amount of cryoprotective material exposed at the surface would serve as a contaminating layer and prevent the visualization of fine details. Recent unexpected freeze-etch results indicated that it would be useful to compare complementary replicas in which one-half of the frozen-fractured specimen would be shadowed and replicated immediately after fracturing whereas the complement would be etched at -98°C for 1 to 10 minutes before being shadowed and replicated.Standard complementary replica holders (Steere, 1973) with hinges removed were used for this study. Specimens consisting of unfixed virus-infected plant tissue infiltrated with 0.05 M phosphate buffer or distilled water were used without cryoprotectant. Some were permitted to settle through gradients to the desired concentrations of different cryoprotectants.

Ca-Histochemistry in slime mold plasmodia

1978 ◽  
Vol 36 (2) ◽  
pp. 130-131
Author(s):  
Ulrich Dierkes
Keyword(s):  
Physarum Polycephalum ◽  
Carbon Coating ◽  
Freeze Drying ◽  
Freeze Fracture ◽  
Freeze Substitution ◽  
Uranyl Acetate ◽  
Slime Mold ◽  
Staining Procedure ◽  
Protoplasmic Streaming ◽  
Intracellular Ca

Calcium is supposed to play an important role in the control of protoplasmic streaming in slime mold plasmodia. The motive force for protoplasmic streaming is generated by the interaction of actin and myosin. This contraction is supposed to be controlled by intracellular Ca-fluxes similar to the triggering system in skeleton muscle. The histochemical localisation of calcium however is problematic because of the possible diffusion artifacts especially in aquous media.To evaluate this problem calcium localisation was studied in small pieces of shock frozen (liquid propane at -189°C) plasmodial strands of Physarum polycephalum, which were further processed with 3 different methods: 1) freeze substitution in ethanol at -75°C, staining in 100% ethanol with 1% uranyl acetate, and embedding in styrene-methacrylate. For comparison the staining procedure was omitted in some preparations. 2)Freeze drying at about -95°C, followed by immersion with 100% ethanol containing 1% uranyl acetate, and embedding. 3) Freeze fracture, carbon coating and SEM investigation at temperatures below -100° C.

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).

Freeze-Fracture Replication in the Ultrastructure of Blue-Green Algae

1967 ◽  
Vol 25 ◽  
pp. 74-75
Author(s):  
L. V. Leak
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Green Algae ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Electron Microscopic ◽  
Photosynthetic Membranes ◽  
Blue Green Algae ◽  
Cell Biol ◽  
Cellular Components

Electron microscopic observations of freeze-fracture replicas of Anabaena cells obtained by the procedures described by Bullivant and Ames (J. Cell Biol., 1966) indicate that the frozen cells are fractured in many different planes. This fracturing or cleaving along various planes allows one to gain a three dimensional relation of the cellular components as a result of such a manipulation. When replicas that are obtained by the freeze-fracture method are observed in the electron microscope, cross fractures of the cell wall and membranes that comprise the photosynthetic lamellae are apparent as demonstrated in Figures 1 & 2.A large portion of the Anabaena cell is composed of undulating layers of cytoplasm that are bounded by unit membranes that comprise the photosynthetic membranes. The adjoining layers of cytoplasm are closely apposed to each other to form the photosynthetic lamellae. Occassionally the adjacent layers of cytoplasm are separated by an interspace that may vary in widths of up to several 100 mu to form intralamellar vesicles.

Freeze fracture imaging and computer simulation of liposome structure: Membrane geometry and defects

1983 ◽  
Vol 41 ◽  
pp. 646-647
Author(s):  
Joseph A. Zasadzinski
Keyword(s):  
Liquid Crystalline ◽  
Freeze Fracture ◽  
Continuum Theory ◽  
Closed Surfaces ◽  
Straight Line ◽  
Low Weight ◽  
Concentric Spheres ◽  
Membrane Geometry ◽  
Dupin Cyclides ◽  
Limiting Case

At low weight fractions, many surfactant and biological amphiphiles form dispersions of lamellar liquid crystalline liposomes in water. Amphiphile molecules tend to align themselves in parallel bilayers which are free to bend. Bilayers must form closed surfaces to separate hydrophobic and hydrophilic domains completely. Continuum theory of liquid crystals requires that the constant spacing of bilayer surfaces be maintained except at singularities of no more than line extent. Maxwell demonstrated that only two types of closed surfaces can satisfy this constraint: concentric spheres and Dupin cyclides. Dupin cyclides (Figure 1) are parallel closed surfaces which have a conjugate ellipse (r1) and hyperbola (r2) as singularities in the bilayer spacing. Any straight line drawn from a point on the ellipse to a point on the hyperbola is normal to every surface it intersects (broken lines in Figure 1). A simple example, and limiting case, is a family of concentric tori (Figure 1b).To distinguish between the allowable arrangements, freeze fracture TEM micrographs of representative biological (L-α phosphotidylcholine: L-α PC) and surfactant (sodium heptylnonyl benzenesulfonate: SHBS)liposomes are compared to mathematically derived sections of Dupin cyclides and concentric spheres.

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