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L H Margaritis

Publications and source records attributed to L H Margaritis.

32 records · Page 2Linked to original sources
PubMed1991

The egg-shell of Drosophila melanogaster. VI, Structural analysis of the wax layer in laid eggs.

Utilizing freeze-fracturing conventional electron microscopy and scanning electron microscopy methods, a wax layer was identified, sealing the oocyte of Drosophila melanogaster. In mature egg-shells wax forms a hydrophobic layer surrounding the oocyte and lying between, and in very close contact with the vitelline membrane (interiorly) and the crystalline intermediate chorionic layer (exteriorly). In cross-fractured views it is less than 50 A thick whereas in longitudinal fracturing it reveals smooth fracture faces of a multilayered material in the form of hydrophobic areas or plaques (0.5-1 microns in diameter) which are partially overlapping and highly compressed between the vitelline membrane and the innermost chorionic layer. The evidence for this layer being a wax are the facts that a) it is not preserved in conventional fat-extracting electron microscopy methods, b) it directs laterally the fracture planes during freeze-fracturing and reveals smooth fracture faces. Analysis of the structural features of wax in mature egg-shell in various species of Drosophilidae have shown that the wax layer exhibits indistinguishable (among the species) hydrophobic plaques, which have the same size and thickness with Drosophila melanogaster. These data provide structural evidence explaining the physiological resistance of the insect eggs studied, against water loss or water uptake, whenever they are laid on substrates with extreme environmental conditions. In addition, the data demonstrate how an extracellular substance can be organized to perform that function.

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PubMed1991

Three-dimensional reconstruction of innermost chorion layer of Drosophila grimshawi and Drosophila melanogaster eggshell mutant fs(1)384.

A low-resolution three-dimensional structure of the crystalline innermost chorionic layer (ICL) of the Hawaiian species Drosophila grimshawi and the Drosophila melanogaster eggshell mutant fs(1)384 has been calculated from electron microscope images of tilted negatively stained specimens. The isolated ICL of Drosophila grimshawi is a three-layer structure, about 36 nm thick, whereas the ICL of Drosophila melanogaster eggshell mutant fs(1)384 is a single layer, about 12 nm thick. Each unit in both crystalline structures includes octamers made up of four heterodimers. Crosslinks between the structural elements, both within and between unit cells form an interconnecting network, apparently important in maintaining the integrity of the layer. A model which may account for the ICL self-assembly formation in vivo and the ICL observed lattice polymorphism is proposed, combining data from the three-dimensional reconstruction work and secondary structure features of the ICL component proteins s36 and s38.

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PubMed1991

A structural protein that plays an enzymatic role in the eggshell of Drosophila melanogaster.

E.S.P. is responsible for the hardening process of the egg-shell at the end of oogenesis (stage 14B) and constitutes a structural component. By immunoblotting, using polyclonal rabbit anti-HRP antibody and anti-rabbit IgG-HRP or Protein A-1251 as second antibody, one major band with MW 38KD on nitrocellulose filter showed positive reaction. We conclude that the E.S.P. is identical to the S38 chorionic protein. Morphological immunogold staining, using pre-embedding procedure, revealed positive reaction in the innermost chorionic layer (ICL) and the endochorion of the eggshell. In addition, electron probe X-ray microanalysis revealed the existence of 37% calcium (explained since the enzyme is Ca2(+)-activated) and 5% iron (explained due to the fact that it is a haemoprotein).

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PubMed1988

Structural and biochemical studies on four sex-linked chorion mutants of Drosophila melanogaster.

Four female-sterile mutants, fs(1)K451, fs(1)K1214, fs(1)K575TS, and fs(1)384, were studied in terms of chorion structure and chorion protein composition. The first three of these mutants cause morphological defects, ie, substantial underproduction and disruption of the endochorion, correlated with underproduction of the six major chorion proteins, s15-s38; the phenotypes are consistent with the observation that these mutants interfere with amplification of the major chorion genes that encode the s15-s38 proteins [Orr et al.: Proc Natl Acad Sci USA 81:3773-3777, 1984; Komitopoulou et al.: Dev Genet 7:75-80, 1986]. The fourth mutant, fs(1)384, and its alleles do not interfere with production of the major chorion proteins and the morphologically detectable bulk of the endochorion but lead to failure of endochorionic organization. Apparently this complementation group is responsible for a minor chorion product, which is evidently important morphogenetically and which is processed posttranslationally in a complex manner [Bauer and Waring: Dev Biol 121:349-358, 1987].

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PubMed1987

The corneal epithelium basement membrane complexes after alkali burn: an ultrastructural study.

Chemical burns were produced using a 4 N NaOH solution in 21 rabbit corneas. The ultrastructural findings were examined under an electron microscope five minutes and 1, 2, 4, 8, 15, and 30 days after the burn. A morphologic disturbance of the basement membrane and its complexes was detected from the first day of the experiment. These findings suggest that the basement membrane plays an important role in the pathogenesis of corneal recurrent erosions and ulcer formation after chemical burns.

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PubMed1984

Three-dimensional reconstruction of innermost chorion layer from Drosophila melanogaster.

A low-resolution three-dimensional structure of the crystalline innermost chorion layer (ICL) has been calculated from electron microscope images of tilted negatively stained crystals. The isolated ICL is a single layer, about 12 nm thick and appears to be made up of two types of subunits, each approximately 3 nm in diameter, arranged regularly as groups of four heterodimers in space group C222. Linking density between these groups of subunits, maintaining the integrity of the layer, appears to be confined mainly to the outer surfaces of the ICL.

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PubMed1980

The eggshell of Drosophila melanogaster. I. Fine structure of the layers and regions of the wild-type eggshell.

The fine structure of the several layers and regional specializations in the Drosophila melanogaster eggshell has been studied by a combination of shell isolation procedures and ultrastructural techniques (conventional TEM, whole-mount TEM, SEM, HVEM, freeze-fracture electron microscopy utilizing rotary replication, shadow casting, optical diffraction and stereo imaging). The main shell consists of 5 layers: the vitelline membrane (300 nm thick), the wax layer, the innermost chorionic layer (40-50 nm), the endochorion (500-700 nm), and the exochorion (300-500 nm). The vitelline membrane consists of irregularly organized particles. The wax layer appears to contain multilayered hydrophobic plates which split tangenitally upon freeze fracturing. The innermost chorionic layer is composed of a crystalling lattice. The endochorion is made of a thin (40 nm) fenestrated floor composed of 40-nm fibres and an outer solid (200 nm) roof covered with a network of 40-nm strands. Intermittently spaced pillar connect these 2 parts. Similarities in the substructure of the floor, pillars and roof suggest that they may be composed of similar or identical structural elements. The specialized regions of the shell are the 2 respiratory appendages, the operculum area and the posterior pole. The appendages exhibit 2 sharply distinct surfaces, a dorsal side with isolated 1.5-micrometer plaques and a ventral side with strands of 40-50 nm connected in a network with openings of 70-80 nm. The operculum area, which includes the micropoyle and the collar, is distinguished by 3 unique types of cell imprints. The posterior pole contains 2 distinctive populations of cell imprints: the central area has very thin intercellular ridges and a thin, perforated, endochorionic roof, while the peripheral area contains mixed, thick and thin, intercellular ridges and serves as a transition zone to the main shell pattern. The pillars in the central area of the posterior pole have a distinct arrangement, forming one peripheral circle within each cell imprint. An analysis utilizing structural and developmental criteria indicates that as many as ten different populations of follicular epithelial cells may be involved in the construction of the various regions of the Drosophila eggshell.

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PubMed1979

Structural and image analysis of a crystalline layer from dipteran eggshell.

A crystalline layer has been identified as a constituent of the eggshell in the dipteran Drosophila melanogaster. This 400A thick intermediate chorionic layer (ICL) is composed of eight 50A thick sublayers and lies between the vitelline membrane and the endochorion. Whole mount views of isolated ICL after negative staining reveal P2 planar periodicity which, when analyzed further by optical diffraction and filtering, showed 1st (100A), 2nd, 3rd and 4th order reflections.

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PubMed1979

Response of the cornea to far ultraviolet light: an ultrastructural study.

Quantitative changes were measured and ultrastructural alterations were detected in rabbit corneas after exposure to a far ultraviolet irradiation (UV). Low dose exposure (105 - 8 x 105 erg/cm2) resulted one day later in a thicker epithelium and loss of the intercellular permeability barrier. In corneas exposed to higher doses (5 x 106 erg/cm2), the epithelium was considerably thinner one day later, whereas 4 days later it was of normal thickness and the endothelial cells became degenerated. The highest UV dose exposure used (a total of 20 x 106 erg/cm2 given in 4 equal daily doses) resulted in (1) swelling of the stroma and disorganization of the collagen fibers; (2) disappearance of the basement membrane; (3) degeneration of the endothelial cells; and (4) considerable reduction of epithelium thickness.

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PubMed1977

Rotary replication for freeze-etching.

Rotary replication has been adapted to freeze-etching and evaluated using T4 polyheads, erythrocyte ghosts, and chloroplast membranes. Conventional electron microscopy, electron diffraction, and optical diffraction and filtering indicate that platinum-carbon rotary replication renders radially symmetrical contrast and 25 A resolution to freeze-etched specimens so as to clarify subunit structure not normally evident in unidirectional shadow replicas.

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PubMed1976

Three dimensional structure of the endochorion in wild type Drosophila melanogaster.

The endochorion of the Drosophila melanogaster eggshell has been isolated and its structure examined by transmission and scanning electron microscopy. In the main shell the thin inner endochorion (floor) shows small pits or pores and characteristic fenestrae crossed by irregular strands; vertical columns surrounded by air spaces separate this layer from the thick outer endochorion (roof) which is covered by a regular, external reticulum of thin fibers. The endochorion of the respiratory horns has sharply divided dorsal and ventral surfaces, with characteristic structures somewhat reminiscent of the floor and the external reticulum of the roof, respectively.

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PubMed1976

Quantitative and comparative ultrastructure of the vertebrate cornea. I. Urodele Amphibia.

The cornea of the urodele amphibian Triturus c. cristatus was studied ultrastructurally in order to provide the basis for a comparison among corneas throughout the vertebrate phylum. The cornea of this salamander consists of relatively thick epithelium and basement membrane and thin Descemet's membrane, unlike the mammalian corneas. The outermost epithelial cells contain Ruthenium Red stainable extracellular filaments and intracellular vesicles which are thought to play a role in the process of lubricating the corneal surface. Occluding junctions have been observed in the apical region of the superficial epithelial cells and are considered as barriers to the intercellular passage of material. A thin substantia propria (stroma) consists of about 40 collagenous highly organized lamellae. The thicknesses of the basement membrane, Descemet's membrane and the epithelium are believed to represent the primitive situation in the process of corneal evolution.

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PubMed1976

Quantitative changes and ultrastructural alterations of the cornea in response to ultraviolet light. II. Effects of amphibia; elucidation of desmosomal structure and basement membrane synthesis.

The effects of far ultraviolet light irradiation upon an amphibian cornea were studied to compare the effects observed both quantitatively and ultrastructurally with data obtained after UV irradiation of mammalian corneas. The ultimate goal of this series of investigations is the elucidation of the alterations and the regeneration mechanisms, which might reflect existing morphological diversities among the species, observed in vertebrate corneas following exposure to UV light. It was found that while the epithelial cells undergo oedema after low dose exposures and are gradually damaged after high doses of UV light, 2-4 days leter a new epithelium has been formed. Intercellular permeability is increased by low dose exposure as was detected by the penetration of Ruthenium Red into the intercellular clefts. Under these conditions desmosomal structure revealed a 21-laminar configuration. The basement membrane of the amphibian, unlike that of the mammal, does not dissolve away upon exposure but shows localized disruptions which are thought to accommodate the passage of leucocytes from stroma to epithelium. That a new basement membrane is subsequently formed is evident by the existence of extracellular and intracellular secretion granules. In comparison to irradiated rabbit corneas, this stroma remains remarkably at the same thickness following a high dose exposure although a noticeable disorganization of collagen arrangement is apparent. Finally, as in the case of the rabbit corneas, a secondary degeneration of endothelium was observed 4 days after a moderate dose exposure.

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