The ultrastructure of the skeleton and skeletogenic tissues of the temperate coral Caryophyllia smithii

1990 ◽  
Vol 70 (2) ◽  
pp. 295-310 ◽  
Author(s):  
Martin D'A.A. Le Tissier
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Organic Matrix ◽  
Secretory Vesicles ◽  
Intercellular Spaces ◽  
Transmission Electron ◽  
Temperate Coral ◽  
Zooxanthellate Coral ◽  
Membrane Bound ◽  
Calicoblastic Ectoderm

The skeleton and calicoblastic ectoderm of the scleractinian non-zooxanthellate coral Caryophyllia smithii were investigated by light microscopy, scanning and transmission electron microscopy. Except for some costal spines, the skeleton was fasciculate. Fasciculi were made up of bundles of crystalline needles, each crystalline needle consisting of a number of linear series of small (<1 μm) rounded crystals. Fractured skeletons showed the fasciculi to be arranged into layers and that within some septa, theca and costal spines there were spaces that contained neither mineral nor organic matter. These spaces could also be found at the growing edges of septa and theca. Demineralization of the skeleton revealed an organic matrix whose configuration mirrored the architecture of the skeleton. In areas of the skeleton where deposition was occurring the overlying calicoblastic ectoderm was relatively thin with prominent intercellular spaces and secretory vesicles. In contrast, over non-depositing areas the calicoblastic ectoderm was thick and contained residual bodies, nematocysts and membrane-bound granules. The results are compared and contrasted with those from scleractinian corals that have endosymbiotic zooxanthellae.

The development of the notochord in the chick embryo, studied by scanning and transmission electron microscopy

Development ◽  
1976 ◽  
Vol 35 (2) ◽  
pp. 383-401
Author(s):  
Mary Bancroft ◽  
Ruth Bellairs
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Chick Embryo ◽  
Intercellular Spaces ◽  
Notochordal Cells ◽  
Fibrillar Material ◽  
Transmission Electron ◽  
Phases Of Development ◽  
Scanning Electron

The notochord of the chick embryo between stages 5 and 23 inclusive has been studied by scanning electron microscopy, supplemented by transmission electron microscopy. Three main phases of development are described, and these have been designated: bilaminar; rodlike, unvacuolated; rod-like and vacuolated. The change in shape of the organ from bilaminar to rod-like is accompanied by changes in the shape, orientation and position of the cells, an increase in the complexity of the cell contacts, and the laying down of a basal lamina. The change from the unvacuolated to the vacuolated phase is accompanied by increasing complexity within the cytoplasm. Most of the vacuoles are intracellular and appear empty though some contain a granular material. The notochordal sheath appears to be secreted by the notochordal cells and fine fibrillar material has been seen in the intercellular spaces. By stage 23, most of the notochordal cells have become so highly vacuolated that the cytoplasm has become closely packed around the nucleus.

scholarly journals The Transport System of Nacre Components through the Surface Membrane of Gastropods

2016 ◽  
Vol 672 ◽  
pp. 103-112 ◽  
Author(s):  
Elena Macías-Sánchez ◽  
Antonio G. Checa ◽  
Marc G. Willinger
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Calcium Carbonate ◽  
Lamellar Structure ◽  
Surface Membrane ◽  
Organic Matrix ◽  
X Ray ◽  
Transmission Electron ◽  
Means Of Transmission ◽  
Electron Energy Loss

The surface membrane is a lamellar structure exclusive of gastropods that is formed during the shell secretion. It protects the surface of the growing nacre and it is located between the mantle epithelium and the mineralization compartment. At the mantle side of the surface membrane numerous vesicles provide material, and at the nacre side, the interlamellar membranes detach from the whole structure. Components of nacre (glycoproteins, polysaccharides and calcium carbonate) cross the structure to reach the mineralization compartment, but the mechanism by which this occurs is still unknown. In this paper we have investigated the ultrastructure of the surface membrane and the associated vesicle layer by means of Transmission Electron Microscopy. Electron Energy Loss Spectroscopy and Energy-dispersive X-ray Spectroscopy were used for elemental analysis. The analyses revealed the concentration of calcium in the studied structures: vesicles, surface membrane, and interlamellar membranes. We discuss the possible linkage of calcium to the organic matrix.

scholarly journals Infection With a Novel Rickettsiella Species in Emperor Scorpions (Pandinus imperator)

2020 ◽  
Vol 57 (6) ◽  
pp. 858-870
Author(s):  
Sushan Han ◽  
Aníbal G. Armién ◽  
Janet E. Hill ◽  
Champika Fernando ◽  
Dan S. Bradway ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Rapid Progression ◽  
Protein Coding ◽  
Hematopoietic Organ ◽  
Transmission Electron ◽  
History Of ◽  
Gross Lesions ◽  
Membrane Bound ◽  
Methenamine Silver

Rickettsiella infection was diagnosed in 4 adult emperor scorpions ( Pandinus imperator) from 2 different collections over a 3-year period. One case had a 2-day history of weakness, failure to lift the tail, or respond to stimulation, with rapid progression to death. The other 3 cases were found dead. There were no gross lesions, but histologically the hemolymphatic vasculature and sinuses, presumed hematopoietic organ, heart, midgut and midgut diverticula, nerves, and skeletal muscle were infiltrated with phagocytic and granular hemocytes with necrosis. Phagocytic hemocytes contained abundant intracellular microorganisms that were Fite’s acid-fast-positive, Macchiavello-positive, variably gram-positive or gram-negative, and Grocott’s methenamine silver-negative. By transmission electron microscopy, hemocytes contained numerous phagocytic vacuoles with small dense bacterial forms (mean 0.603 × 0.163 μm) interspersed with large bacterial forms (mean 1.265 × 0.505 μm) and few intermediary forms with electron-dense nucleoids and membrane-bound crystalline arrays (average 4.72 μm). Transmission electron microscopy findings were consistent with bacteria of the family Coxiellaceae. Based on sequencing the 16S ribosomal RNA gene, the identity was confirmed as Rickettsiella, and phylogenetic analysis of protein-coding genes gidA, rspA, and sucB genes suggested the emperor scorpion pathogen as a new species. This study identifies a novel Rickettsiella causing infection in emperor scorpions and characterizes the unique pathological findings of this disease. We suggest this organism be provisionally named Rickettsiella scorpionisepticum.

Phenethyl Isothiocyanate (PEITC) Regulates Autophagy in Chronic Lymphocytic Leukemia.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2906-2906
Author(s):  
Jemimah Adams ◽  
R Gitendra Wickremasinghe ◽  
Archibald G Prentice ◽  
Jonathan C. Strefford ◽  
Andrew Duncombe ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Annexin V ◽  
Lymphocytic Leukemia ◽  
Phenethyl Isothiocyanate ◽  
Autophagosome Formation ◽  
Double Membrane ◽  
Transmission Electron ◽  
Membrane Bound ◽  
Autophagy Pathway

Abstract Abstract 2906 Chronic Lymphocytic leukemia (CLL) is currently incurable using conventional therapies. CLL cells can evade killing by various therapeutic strategies. However the precise mechanisms are currently unknown. Autophagy is regulated by a complex system of proteins, and is used by both normal and malignant cells as a protective mechanism against cellular stress induced by starvation, hypoxia, reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress. In malignant cells autophagy was shown to promote tumorigenesis and/or resistance to chemotherapy. Therefore we hypothesized that autophagy may play a role in CLL biology. Autophagy can also promote cell death when stress signals are elevated above a particular threshold for a prolonged period of time. In this study we investigated the basal expression levels of autophagy specific genes and the effect of autophagy specific inhibitors (Bafilomycin, 3-methyladenine and hydroxychloroquine) and inducers (Phenethyl isothiocyanate) on CLL survival. Phenethyl isothiocyanate (PEITC) is about to enter clinical trials for CLL (NCT00968461). We have investigated induction of components of the autophagic pathway following treatment of CLL cells in vitro with a range of chemical inhibitors. Immunoblotting was carried out to investigate components of the autophagy pathway using phosphorylation state-specific and pan-reactive antibodies. Bafilomycin (BAF), 3-methyladenine (3-MA) and hydroxychloroquine (HCQ) toxicity towards CLL samples were evaluated by Annexin V/PI staining, MTT assay and immunoblotting for cleavage of the caspase 3 substrate poly(ADP ribose) polymerase (PARP) from its 116KDa to its 85KDa form. PEITC was used at concentrations between 2.5 and 25μM to investigate its effect on signaling. Autophagy was quantitated by immunoblotting of LC3-I and LC3-II. Lipidation of LC3 from LC3-I to LC3-II is a surrogate marker of autophagy and is essential for autophagasome formation. Immunoblotting was also performed for ATG3, ATG5 and ATG7, key components of the autophagy pathway. Monodansylcadaverine (MDC) was used with immunofluorescence and FACS analysis to investigate increases in autophagasome formation. Transmission electron microscopy (TEM) was used to confirm double membrane bound autophagosomes. Co-immunoprecipitation was used to evaluate if Beclin-1 was sequestered by Bcl-2 preventing autophagy. Its release from Bcl-2 enables Beclin-1 to interact with other autophagy specific proteins and initiates autophagasome formation. LC3-I was lipidated to LC3-II (p=0.019) and ATG3 (p=0.021) was upregulated to a greater extent in CLL samples compared with normal B-cell controls at basal levels. This suggested that autophagy was active to a greater extent in CLL samples compared with normal individuals. In addition Beclin was dissociated from Bcl-2 in CLL samples indicating that autophagy was active. Autophagy appears to be a pro-survival mechanism in untreated CLL cells as inhibiting basal levels of autophagy with autophagy inhibitors BAF (50–200nM), 3-MA (5–10mM) and hydroxychlorquine (5–10μM) resulted in CLL apoptosis as shown by MTT, Annexin V/PI analysis and PARP cleavage. Interestingly augmenting autophagy was also capable of inducing apoptosis in CLL samples. Treatment with PEITC caused an increase in punctate staining using MDC which is suggestive of autophagosome formation. We went on to determine that PEITC further induced LC3-II lipidation using immunoblotting and showed a substantial increase in overall LC3 protein expression. PEITC also induced the expression of ATG3, a key protein in the autophagy pathway. We then evaluated autophagosome formation using TEM (Figure 1). Our data showed greater numbers of autophagosomes in the PEITC treated samples compared to the untreated controls. Therefore autophagy in CLL sits on a knife-edge, such that perturbations that either increase pro- death or decrease pro-survival autophagy signals can result in CLL cell death, depending on the duration and intensity of the signal. Figure 1. Transmission electron microscopy of CLL cells CLL cells were treated with 10μM PEITC. Double membrane bound organelles were found in the CLL cells after treatment which were not present in the no addition control (depicted by the arrows). These organelles are autophagsomes. Magnification (left picture) ruler is 500nM, (right picture) ruler is 100nM Figure 1. Transmission electron microscopy of CLL cells . / CLL cells were treated with 10μM PEITC. Double membrane bound organelles were found in the CLL cells after treatment which were not present in the no addition control (depicted by the arrows). These organelles are autophagsomes. Magnification (left picture) ruler is 500nM, (right picture) ruler is 100nM Disclosures: No relevant conflicts of interest to declare.

Matrix-Mediated Biomineralization in Marine Mollusks: A Combined Transmission Electron Microscopy and Focused Ion Beam Approach

2011 ◽  
Vol 17 (2) ◽  
pp. 220-225 ◽  
Author(s):  
Martin Saunders ◽  
Charlie Kong ◽  
Jeremy A. Shaw ◽  
Peta L. Clode
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
High Strength ◽  
Organic Matrix ◽  
Dark Field ◽  
Transmission Electron ◽  
Annular Dark Field ◽  
The Relationship

AbstractThe teeth of the marine mollusk Acanthopleura hirtosa are an excellent example of a complex, organic, matrix-mediated biomineral, with the fully mineralized teeth comprising layers of iron oxide and iron oxyhydroxide minerals around a calcium apatite core. To investigate the relationship between the various mineral layers and the organic matrix fibers on which they grew, sections have been prepared from specific features in the teeth at controlled orientations using focused ion beam processing. Compositional and microstructural details of heterophase interfaces, and the fate of the organic matrix fibers within the mineral layers, can then be analyzed by a range of transmission electron microscopy (TEM) techniques. Energy-filtered TEM highlights the interlocking nature of the various mineral phases, while high-angle annular dark-field scanning TEM imaging demonstrates that the organic matrix continues to exist in the fully mineralized teeth. These new insights into the structure of this complex biomaterial are an important step in understanding the relationship between its structural and physical properties and may help explain its high strength and crack-resistance behavior.

scholarly journals FRTL-5 Rat Thyroid Cells Release Thyroglobulin Sequestered in Exosomes: A Possible Novel Mechanism for Thyroglobulin Processing in the Thyroid

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Pavel Vlasov ◽  
Sonia Q. Doi ◽  
Donald F. Sellitti
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Western Blotting ◽  
Thyroid Stimulating Hormone ◽  
Cellular Proteins ◽  
Thyroid Cells ◽  
Normal Thyroid ◽  
Transmission Electron ◽  
Membrane Bound ◽  
Rat Thyroid

Exosomes are 30–100 nm, membrane-bound vesicles containing specific cellular proteins, mRNAs, and microRNAs that take part in intercellular communication between cells. A possible role for exosomes in thyroid function has not been fully explored. In the present study, FRTL-5 rat thyroid cells were grown to confluence and received medium containing either thyroid stimulating hormone (TSH), exogenous bovine thyroglobulin (bTg), or neither additive for 24 or 48 hours followed by collection of spent medium and ultracentrifugation to isolate small vesicles. Transmission electron microscopy and Western blotting for CD9 indicated the presence of exosomes. Western blotting of exosome extract using a monoclonal anti-Tg antibody revealed a Tg-positive band at ~330 kDa (the expected size of monomeric Tg) with a higher density in TSH-treated cells compared to that in untreated cells. These results are the first to show that normal thyroid cells in culture produce exosomes containing undegraded Tg.

Ultrastructural Changes in Root Cap Cells of Two Australian Native Grass Species Following Exposure to Aluminium

1997 ◽  
Vol 24 (2) ◽  
pp. 165 ◽  
Author(s):  
Simon A. Crawford ◽  
Sabine Wilkens
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Root Cap ◽  
Ultrastructural Changes ◽  
Grass Species ◽  
Aluminium Tolerance ◽  
Secretory Vesicles ◽  
Native Grass ◽  
Transmission Electron

Transmission electron microscopy was used to investigate ultrastructural changes in root cap cells of two aluminium-tolerant native grass species, Danthonia linkii Kunth and Microlaena stipoides (Labill.) R.Br., following exposure to Al. Quantitative differences in root cap cells and organelles in response to 0–10 ppm aluminium were determined using image analysis of electron micrographs. Changes to the size of root cap cells due to exposure to Al were similar in the two species with a low Al concentration (1–2 ppm) resulting in larger cells, while higher Al levels (5–10 ppm) reduced cell size. In peripheral cap cells of the more Al-tolerant M. stipoides, the size of secretory vesicles was not affected by exposure to Al, while in peripheral cap cells of the less Al-tolerant D. linkii, exposure to Al resulted in significantly smaller secretory vesicles being produced. Central root cap cells from control plants of M. stipoides contained 90% more dictyosomes and had 50% larger amyloplasts than D. linkii. Measurement of mucilage droplets showed that roots of M. stipoides produced much more mucilage than D. linkii. Exposure of roots of M. stipoides to Al in the range 2–10 ppm had no effect on the size of mucilage droplets produced. The possible role of mucilage production in aluminium tolerance is discussed.

In situ transmission electron microscopy observation of reversible deformation in nacre organic matrix

2008 ◽  
Vol 23 (5) ◽  
pp. 1466-1471 ◽  
Author(s):  
Taro Sumitomo ◽  
Hideki Kakisawa ◽  
Yusuke Owaki ◽  
Yutaka Kagawa
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Mechanical Performance ◽  
Protein Structures ◽  
Organic Matrix ◽  
Organic Polymer ◽  
Transmission Electron ◽  
Strong Adhesion ◽  
Abalone Shell

The deformation behavior of the organic polymer matrix of the biocomposite nacre structure in abalone shell was investigated by in situ straining during transmission electron microscopy (TEM). We observed strong adhesion to mineral plates and high ductility of the organic matrix, confirming a crack-bridging toughening mechanism. In addition, direct observation of reversible mechanical behavior was made in the viscoelastic reformation of matrix ligaments after failure. Crystalline β-sheet structures identified through electron diffraction suggested the presence of protein structures similar to spider or cocoon silk, and the reversible mechanism was attributed to hydration-induced unfolding and refolding of domains in these silklike proteins. This work provides further insight into the molecular and nanoscale behavior of nacre organic matrix and its contribution to bulk mechanical performance.

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