Surface membrane regeneration in deciliated Tetrahymena

1983 ◽  
Vol 62 (1) ◽  
pp. 407-417
Author(s):  
N.E. Williams
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Membrane Proteins ◽  
Cell Surface ◽  
Surface Membrane ◽  
Membrane System ◽  
Ciliated Protozoa ◽  
Membrane Flow ◽  
Transmission Electron ◽  
Surface Marking

The induced synthesis of identified surface membrane proteins has been demonstrated in deciliated Tetrahymena. Cells in the process of regenerating cilia were also studied using transmission electron microscopy in order to obtain information on the deployment of new membrane at the cell surface. The results obtained suggest a pattern of membrane flow that includes the ‘pellicular alveoli’, a subsurface membrane system characteristically present in ciliated protozoa. The results of 125I surface-marking experiments were consistent with the notion that new membrane is added initially in non-ciliated regions, then subsequently flows laterally to cover regenerating cilia.

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.

Transmission electron microscopy (TEM) of equine conceptuses at 14 and 16 days of gestation

2010 ◽  
Vol 22 (2) ◽  
pp. 405 ◽  
Author(s):  
Ingrid Walter ◽  
Waltraud Tschulenk ◽  
Sven Budik ◽  
Christine Aurich
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Surface ◽  
Specific Cell ◽  
Fluid Uptake ◽  
Embryonic Disc ◽  
Transmission Electron ◽  
Cellular Junctions ◽  
Endodermal Cells ◽  
Specific Cell Surface

The present study gives a detailed ultrastructural description of equine conceptuses at Day 14 (n = 2) and Day 16 (n = 3) after ovulation. Whereas on Day 14 only primitive structures were seen, on Day 16 neurulation and formation of mesodermal somites had taken place. The ectoderm of the embryo itself and the surrounding trophoblast ectodermal cells were characterised by specific cell surface differentiations. At the embryonic ectodermal cell surface (14 and 16 days) remarkable protruded and fused cytoplasmic projections were seen, typically associated with macropinocytotic events involved in macromolecule and fluid uptake. This finding adds an important point to the expansion mode of the hypotone equine conceptus, which is characterised by ‘uphill’ fluid uptake. Numerous microvilli and coated endocytotic pits at the apical trophoblast membrane emphasised its absorptive character. Endodermal cells were arranged loosely with only apically located cellular junctions leaving large intercellular compartments. At the border of the embryonic disc apoptotic cells were regularly observed indicating high remodelling activities in this area. Conspicuous blister-like structures between ectoderm and mesoderm were seen in the trilaminar part of Day-14 and -16 conceptuses. These were strictly circumscribed despite not being sealed by cellular junctions between germinal layers. It is possible that these blisters are involved in embryo positioning; however, further studies are needed to verify this.

scholarly journals Storage pool deficiency in cattle with the Chediak-Higashi syndrome results from an absence of dense granule precursors in their megakaryocytes

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1726-1734 ◽  
Author(s):  
M Menard ◽  
KM Meyers
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Transmission Electron Microscopy ◽  
Membrane System ◽  
Dense Granule ◽  
Virtual Absence ◽  
Dense Granules ◽  
Storage Pool ◽  
Transmission Electron ◽  
Chediak Higashi Syndrome

Abstract Platelets from cattle with the Chediak-Higashi syndrome (CHS) have a storage pool deficiency and virtual absence of platelet dense granules. Megakaryocytes (MKs) from five control (n = 135) and five CHS (n = 133) cattle were evaluated using standard transmission electron microscopy. Osmiophilic dense granules were not observed in control or CHS MKs. In MKs from normal cattle, clear vesicles of 200- to 650-nm diameter bounded by a sharp membrane were observed. They were easily differentiated from the demarcation membrane system, endoplasmic reticulum, and alpha granules. The clear vesicles were virtually absent in MKs from CHS cattle at all stages of maturation. MKs in bone marrow samples from two control (n = 91) and two CHS (n = 61) cattle that had been processed for the uranaffin reaction were also evaluated. The clear vesicles were replaced by uranaffin-positive granules in MKs from control cattle, but positive uranaffin granules were not observed in CHS MKs. These findings indicate that the platelet dense granule storage pool deficiency in CHS cattle results from an anatomic absence of dense granule precursors in maturing and mature CHS MKs.

The translaminal fibrils of the human amnion basement membrane

1989 ◽  
Vol 94 (2) ◽  
pp. 307-318
Author(s):  
S. Campbell ◽  
T.D. Allen ◽  
B.B. Moser ◽  
J.D. Aplin
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Surface ◽  
Basal Lamina ◽  
Ammonium Hydroxide ◽  
Apical Surface ◽  
Lamina Densa ◽  
Transmission Electron ◽  
Extraction Pattern ◽  
Stromal Matrix

The organisation of extracellular matrix beneath the human amniotic epithelium was investigated in order that the co-ordinate synthesis of basal lamina and stroma by these cells could be better understood. Transmission electron microscopy of intact tissue confirmed that stromal matrix fibrils are located between the cell surface and the basal lamina, and also penetrate the lamina. The distribution of the supralaminal fibrils and their association with the lamina was further investigated by scanning electron microscopy (SEM) after removal of the overlying epithelium. Five complementary procedures were used to remove the cells from the underlying lamina. Trypsin-EDTA treatment caused the epithelial cells to retract or detach from the lamina. SDS or ammonium hydroxide was used to extract the epithelium, which was then removed by physical shearing. Transmission electron microscopy (TEM) confirmed that the lamina densa and supralaminal fibres were present after extraction by these agents. Incubation in CHAPS, a zwiterionic detergent, did not remove the epithelium but permitted exposure of the basal lamina by mechanical scoring. Extraction with boric acid followed by osmium tetroxide produced epithelial disruption and revealed the lamina and stroma in different areas. Although the extraction pattern was different in each case, all of the five methods confirmed that individual fibrils and fibril bundles are present on the apical surface of, and enter, the lamina densa. Examination of the stromal surface of the basal lamina after fracture revealed fibrils passing from the stroma into the lamina densa. We therefore suggest that, in this tissue, newly synthesised stromal matrix components appear in an assembled fibrillar form between the basal cell surface and the basal lamina before becoming associated with the sublaminal stroma.

scholarly journals Clostridium perfringens produces an adhesive pilus required for the pathogenesis of necrotic enteritis in poultry

2021 ◽  
Author(s):  
D. Lepp ◽  
Y. Zhou ◽  
S. Ojha ◽  
I. Mehdizadeh Gohari ◽  
J. Carere ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Surface ◽  
Clostridium Perfringens ◽  
Direct Evidence ◽  
Necrotic Enteritis ◽  
Null Mutant ◽  
Transmission Electron ◽  
Mutant Strains ◽  
Important Disease

Clostridium perfringens Type G strains cause necrotic enteritis (NE) in poultry, an economically important disease that is a major target of in-feed antibiotics. NE is a multifactorial disease, involving not only the critically-important NetB toxin, but also additional virulence and virulence-associated factors. We previously identified a C. perfringens chromosomal locus (VR-10B) associated with disease-causing strains that is predicted to encode a sortase-dependant pilus. In the current study, we sought to provide direct evidence for the production of a pilus by C. perfringens and establish its role in NE pathogenesis. Pilus structures in virulent C. perfringens strain CP1 were visualized by transmission electron microscopy (TEM) of immuno-gold labelled cells. Filamentous structures were observed extending from the cell surface in wild-type CP1, but not from isogenic pilin-null mutant strains. In addition, immuno-blotting of cell surface proteins demonstrated that CP1, but not the null mutant strains, produced a high molecular weight ladder-like pattern characteristic of a pilus polymer. Binding to collagen types I, II and IV was significantly reduced (Tukey’s; p<0.01) in all three pilin mutants compared to CP1, and could be specifically blocked by CnaA and FimA antisera, indicating that these pilins participate in adherence. Furthermore, both fimA and fimB null mutants were both severely attenuated in their ability to cause disease in an in vivo chicken NE challenge model. Together, these results provide the first direct evidence for the production of a sortase-dependant pilus by C. perfringens, and confirm its critical role in NE pathogenesis and collagen-binding. Importance In necrotic enteritis (NE), an intestinal disease of chickens, Clostridium perfringens cells adhere tightly to damaged intestinal tissue, but the factors involved are not known. We previously discovered a cluster of C. perfringens genes predicted to encode a pilus, a hair-like bacterial surface structure commonly involved in adherence. In the current study, we have directly imaged this pilus using transmission electron microscopy (TEM). We also show that inactivation of the pilus genes stops pilus production, significantly reducing the bacterium's ability to bind collagen and cause disease. Importantly, this is the first direct evidence for the production of a sortase-dependant pilus by C. perfringens, revealing a promising new target for developing therapeutics to combat this economically important disease.

Insights into the mechanism of high lipid–detergent crystallization of membrane proteins

2021 ◽  
Vol 54 (6) ◽  
Author(s):  
Sofia Trampari ◽  
Caroline Neumann ◽  
Samuel J. Hjorth-Jensen ◽  
Azadeh Shahsavar ◽  
Esben M. Quistgaard ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Phase Diagram ◽  
Transmission Electron Microscopy ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Detergent Concentration ◽  
Transmission Electron ◽  
High Lipid ◽  
High Concentrations

Obtaining well diffracting crystals of membrane proteins is often challenging, but chances can be improved by crystallizing them in lipidic conditions that mimic their natural membrane environments. One approach is the high lipid–detergent (HiLiDe) method, which works by mixing the target protein with high concentrations of lipid and detergent prior to crystallization. Although this approach is convenient and flexible, understanding the effects of systematically varying lipid/detergent ratios and a characterization of the lipid phases that form during crystallization would be useful. Here, a HiLiDe phase diagram is reported for the model membrane protein MhsT, which tracks the precipitation and crystallization zones as a function of lipid and detergent concentrations, and is augmented with data on crystal sizes and diffraction properties. Additionally, the crystallization of SERCA1a solubilized directly with native lipids is characterized as a function of detergent concentration. Finally, HiLiDe crystallization drops are analysed with transmission electron microscopy, which among other features reveals liposomes, stacked lamellae that may represent crystal precursors, and mature crystals with clearly discernible packing arrangements. The results emphasize the significance of optimizing lipid/detergent ratios over broad ranges and provide insights into the mechanism of HiLiDe crystallization.

scholarly journals Disappearance of macrophage surface folds after antibody-dependent phagocytosis.

1981 ◽  
Vol 89 (2) ◽  
pp. 223-229 ◽  
Author(s):  
H R Petty ◽  
D G Hafeman ◽  
H M McConnell
Keyword(s):  
Electron Microscopy ◽  
Cell Surface ◽  
Guinea Pig ◽  
Surface Area ◽  
Surface Membrane ◽  
Peritoneal Macrophages ◽  
Injection Technique ◽  
Surface Areas ◽  
Transmission Electron ◽  
Uptake Data

We have employed the method of Burwen and Satir (J. Cell Biol., 1977, 74:690) to measure the disappearance of surface folds from resident guinea pig peritoneal macrophages after antibody-dependent phagocytosis. Unilamellar phospholipid vesicles containing dimyristoylphosphatidylcholine and 1 mol % dinitrophenyl-epsilon-aminocaproyl-phosphatidylethanolamine, a lipid that possesses a hapten headgroup, were prepared by an ether injection technique. These vesicles were taken up by macrophages in a time- and temperature-dependent fashion. Vesicles that contained ferritin trapped in the internal aqueous volume were identified within macrophages by transmission electron microscopy. Scanning electron microscopy has shown that macrophage surface folds decrease dramatically after phagocytosis. The surface fold length (micrometer) per unit smooth sphere surface area (micrometer2) decreases from 1.3 +/- 0.3 micrometer-1 to 0.53 +/- 0.25 micrometer-1 when cells are incubated in the presence of specific anti-DNP antibody and vesicles at 37 degrees C. No significant effect was observed in the presence of antibody only or vesicles only. Our studies shown that phagocytosis is associated with a loss of cell surface folds and a loss of cell surface area, which is consonant with current views of the endocytic process. On the basis of our uptake data, we estimate that approximately 400 micrometer2 of vesicle surface membrane is internalized. The guinea pig macrophage plasma membrane has a total area of approximately 400 micrometer2 in control studies, whereas the cells have roughly 300 micrometer2 after phagocytosis. These estimates of surface areas include membrane ruffles and changes directly related to changes in cell volume. We suggest that during antibody-dependent phagocytosis a membrane reservoir is made available to the cell surface.

scholarly journals 3D imaging and quantitative analysis of small solubilized membrane proteins and their complexes by transmission electron microscopy

Microscopy ◽  
2012 ◽  
Vol 62 (1) ◽  
pp. 95-107 ◽  
Author(s):  
Ardeschir Vahedi-Faridi ◽  
Beata Jastrzebska ◽  
Krzysztof Palczewski ◽  
Andreas Engel
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Quantitative Analysis ◽  
Membrane Proteins ◽  
3D Imaging ◽  
Transmission Electron

scholarly journals Embedding cell monolayers to investigate nanoparticle-plasmalemma interactions at transmission electron microscopy

2019 ◽  
Vol 63 (1) ◽  
Author(s):  
Manuela Costanzo ◽  
Manuela Malatesta
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Surface ◽  
Inexpensive Method ◽  
Biological Barrier ◽  
Cell Monolayers ◽  
Ultrastructural Evidence ◽  
Transmission Electron ◽  
Uptake Process ◽  
Intracellular Fate

Transmission electron microscopy is the technique of choice to visualize the spatial relationships between nanoconstructs and cells and especially to monitor the uptake process of nanomaterials. It is therefore crucial that the cell surface be preserved in its integrity, to obtain reliable ultrastructural evidence: the plasmalemma represents the biological barrier the nanomaterials have to cross, and the mode of membrane-nanoconstruct interaction is responsible for the intracellular fate of the nanomaterials. In this paper, we describe a simple and inexpensive method to process cell monolayers for ultrastructural morphology and immunocytochemistry, ensuring consistent preservation of the cell surface and of the occurring interactions with nanoparticles of different chemical composition.

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