scholarly journals Vesicle accumulation and exocytosis at sites of plasma membrane disruption.

1995 ◽  
Vol 131 (6) ◽  
pp. 1737-1745 ◽  
Author(s):  
K Miyake ◽  
P L McNeil
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Membrane Disruption ◽  
Cam Kinase ◽  
Forward Scatter ◽  
Quantitative Evidence ◽  
Transmission Electron ◽  
Large Numbers ◽  
Direct Support ◽  
Membrane Compartments

Plasma membrane disruptions are resealed by an active molecular mechanism thought to be composed, in part, of kinesin, CaM kinase, snap-25, and synaptobrevin. We have used HRP to mark the cytoplasmic site of a mechanically induced plasma membrane disruption. Transmission electron microscopy revealed that vesicles of a variety of sizes rapidly (s) accumulate in large numbers within the cytoplasm surrounding the disruption site and that microvilli-like surface projections overlie this region. Scanning electron microscopy confirmed that tufts of microvilli rapidly appear on wounded cells. Three assays, employing the membrane specific dye FM1-43, provide quantitative evidence that disruption induces Ca(2+)-dependent exocytosis involving one or more of the endosomal/lysosomal compartments. Confocal microscopy revealed the presence in wounded cells of cortical domains that were strikingly depleted of FM dye fluorescence, suggesting that a local bolus of exocytosis is induced by wounding rather than global exocytosis. Finally, flow cytometry recorded a disruption-induced increase in cell forward scatter, suggesting that cell size increases after injury. These results provide the first direct support for the hypothesis that one or more internal membrane compartments accumulate at the disruption site and fuse there with the plasma membrane, resulting in the local addition of membrane to the surface of the mechanically wounded cell.

Ultrastructural aspects of coated vesicles in tobacco protoplasts

1981 ◽  
Vol 59 (7) ◽  
pp. 1307-1313 ◽  
Author(s):  
P. van der Valk ◽  
L. C. Fowke
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Plasma Membrane ◽  
Coated Vesicles ◽  
Thin Sections ◽  
Tobacco Protoplasts ◽  
Vesicle Membranes ◽  
Transmission Electron ◽  
Large Numbers ◽  
Inner Surface

The ultrastructure and distribution of coated vesicles in isolated tobacco protoplasts were investigated using transmission electron microscopy of thin sections of whole protoplasts and stained plasma membrane preparations obtained by osmotic bursting of protoplasts attached to coated microscope grids. Large numbers of coated vesicles were associated with both the plasma membrane and the maturing face of dictyosomes. Dictyosome associated coated vesicles were smaller and had less distinct coats and vesicle membranes than those associated with the plasma membrane. Honeycomb structures believed to be aggregations of coats were also associated with the inner surface of the plasma membrane. Our data suggest that coated vesicles are produced by the Golgi apparatus, fuse with the plasma membrane, their coats remaining attached, at least temporarily, to the plasma membrane inner surface.

scholarly journals A novel exosome biogenesis mechanism: multivesicular structures budding and rupturing at the plasma membrane

2019 ◽  
Author(s):  
Stephanie Leon Quinonez ◽  
Ian R. Brown ◽  
Helen E. Grimsley ◽  
Jindrich Cinatl ◽  
Martin Michaelis ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Membrane Association ◽  
Dense Layer ◽  
Physiological Processes ◽  
Exosome Biogenesis ◽  
Transmission Electron ◽  
Typical Morphology ◽  
Intercellular Signalling ◽  
Microscopy Images

AbstractExosomes are small vesicles secreted by the cells, which mediate intercellular signalling and systemic physiological processes. Exosomes are known to originate from the intraluminal vesicles of the multivesicular endosome that fuses with the plasma membrane. We found that the non-small cell lung cancer (NSCLC) cell lines, HCC15 and A549, secreted exosomes with typical morphology and protein contents. Unexpectedly, transmission electron microscopy images indicated that the cells formed multivesicular structures that protruded from the plasma membrane and ruptured to release the exosomes. There were smooth multivesicular structures surrounded by an ordinary looking membrane, multivesicular structures coated by an electron dense layer with regular spacing pattern, and intermediate forms that combined elements of both. Electron microscopy images suggested that exosomes are release from these structures by burst events and not by the conventional fusion process. The molecular details of this novel mechanism for membrane association, deformation and fusion is to be unveiled in the future.

Morphology and ultrastructure of oral strains of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus

1980 ◽  
Vol 30 (2) ◽  
pp. 588-600
Author(s):  
S C Holt ◽  
A C Tanner ◽  
S S Socransky
Keyword(s):  
Electron Microscopy ◽  
Ruthenium Red ◽  
Actinobacillus Actinomycetemcomitans ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Large Numbers ◽  
Haemophilus Aphrophilus ◽  
Ruthenium Red Staining ◽  
Amorphous Surface ◽  
Scanning Electron

Selected human oral and nonoral strains of the genera Actinobacillus and Haemophilus were examined by transmission and scanning electron microscopy. The strains examined were morphologically identical to recognized Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus. By transmission electron microscopy, the cells were typically gram negative in morphology, with several strains possessing some extracellular ruthenium red-staining polymeric material. Numerous vesicular structures, morphologically identical to lipopolysaccharide vesicles, were seen to originate from and be continuous with the surface of the outer membrane. Large numbers of these vesicles were also found in the external environment. Scanning electron microscopic observations revealed that both actinobacilli and haemophili possessed surface projections and an amorphous surface material which connected and covered adjacent cells.

Differential binding of gold-labeled zona pellucida glycoproteins mZP2 and mZP3 to mouse sperm membrane compartments

Development ◽  
1991 ◽  
Vol 113 (1) ◽  
pp. 141-149 ◽  
Author(s):  
S. Mortillo ◽  
P.M. Wassarman
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Transmission Electron ◽  
Acrosomal Membrane ◽  
Membrane Compartments ◽  
Sperm Membrane ◽  
Differential Binding ◽  
Inner Acrosomal Membrane ◽  
Low Levels ◽  
Species Specific

Egg zona pellucida glycoproteins mZP3 and mZP2 serve as primary and secondary sperm receptors, respectively, during initial stages of fertilization in mice [Wassarman (1988) A. Rev. Biochem. 57, 415–442]. These receptors interact with complementary egg-binding proteins (EBPs) located on the sperm surface to support species-specific gamete adhesion. Results of whole-mount autoradiographic experiments suggest that purified egg mZP3 and mZP2 bind preferentially to acrosome-intact (AI) and acrosome-reacted (AR) sperm heads, respectively [Bleil and Wassarman (1986) J. Cell Biol. 102, 1363–1371]. Here, we used purified egg mZP2, egg mZP3 and fetuin, which were coupled directly to colloidal gold (‘gold-probes’), to examine binding of these glycoproteins to membrane compartments of AI and AR sperm by transmission electron microscopy. mZP3 gold-probes were found associated primarily with plasma membrane overlying the acrosomal and post-acrosomal regions of AI sperm heads. They were also found associated with plasma membrane overlying the post-acrosomal region of AR sperm heads. mZP2 gold-probes were found associated primarily with inner acrosomal membrane of AR sperm heads, although some gold was associated with outer acrosomal membrane of AI sperm that had holes in plasma membrane overlying the acrosome. Fetuin gold-probes, used to assess background levels of binding, were bound at relatively low levels to plasma membrane and inner acrosomal membrane of AI and AR sperm, respectively. None of the gold-probes exhibited significant binding to sperm tails, or to red blood cells and residual bodies present in sperm preparations. These results provide further evidence that mZP2 and mZP3 bind preferentially to heads of AR and AI sperm, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

The comparative ultrastructure of spermatozoa from Bothrimonus sturionis Duv. 1842 (Pseudophyllidea), Pseudanthobothrium hanseni Baer, 1956 (Tetraphyllidea), and Monoecocestus americanus Stiles, 1895 (Cyclophyllidea)

1984 ◽  
Vol 62 (6) ◽  
pp. 1059-1066 ◽  
Author(s):  
Barbara M. MacKinnon ◽  
Michael D. B. Burt
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Plasma Membrane ◽  
Main Body ◽  
Posterior Portion ◽  
Transmission Electron ◽  
Dense Nucleus ◽  
Comparative Ultrastructure ◽  
Electron Lucent ◽  
Mature Spermatozoa

The mature spermatozoa from Bothrimonus sturionis (Pseudophyllidea), Pseudanthobothrium hanseni (Tetraphyllidea), and Monoecocestus americanus (Cyclophyllidea) were examined using transmission electron microscopy. Transverse sections of the sperm of B. sturionis indicate that the number of sperm axonemes varies from one to eight, with approximately one-third of the sperm containing two axonemes. Likewise, the number of peripheral microtubules lying just within the external plasma membrane varies from 12 to 20. The nucleus is electron lucent and fibrous in appearance. The spermatozoa of B. sturionis show great variation in the material examined and the majority of them are believed to be aberrant. The spermatozoon of P. hanseni contains a single axoneme with the nucleus wrapped in a crescent around it in the anterior region of the sperm. The posterior portion of the spermatozoon is characterized by a helical flange which projects from the main body of the sperm. The spermatozoon of M. americanus is elongate and slender, containing a single axoneme with an electron-dense nucleus coiled around it in the anterior one-third of the sperm. Electron-opaque bodies, which may be glycogen, fill the cytoplasm. The spermatozoa of all three species contain neither an acrosome nor mitochondria. The flagella of all the spermatozoa have a 9 + "1" arrangement of microtubules. The importance of the ultrastructure of spermatozoa in the phylogeny and taxonomy of cestodes is discussed.

Condensed tannins in sainfoin. 1. A histological and cytological survey of plant tissues

1993 ◽  
Vol 71 (9) ◽  
pp. 1147-1152 ◽  
Author(s):  
G. L. Lees ◽  
N. H. Suttill ◽  
M. Y. Gruber
Keyword(s):  
Electron Microscopy ◽  
Condensed Tannins ◽  
Condensed Tannin ◽  
Plant Tissues ◽  
Onobrychis Viciifolia ◽  
Ultrastructural Studies ◽  
Transmission Electron ◽  
Large Numbers ◽  
Fine Localization ◽  
Cellular Organelles

Sainfoin (Onobrychis viciifolia Scop.) organs and tissues were surveyed for the presence of proanthocyanidins or condensed tannin (CT) in an attempt to learn more about CT formation. Three histological methods were used: a vanillin–HCl stain to localize areas of CT formation through a distinctive cherry-red coloration of CT-containing cells; osmium fixation to further localize CT formation in cellular organelles using light microscopy; and fine localization and examination using electron microscopy. Cells containing CTs were found in relatively large numbers throughout the sainfoin plant in all organs except the roots. No CTs were found in the cotyledons or in some petals. Ultrastructural studies revealed that CTs are restricted to the vacuoles and appear to accumulate on the inner periphery adjacent to the tonoplast or in the centre of the organelle, possibly next to vacuolar invaginations. Key words: condensed tannin, proanthocyanidin, sainfoin, histology, transmission electron microscopy.

The fine structure of Thioploca araucae and Thioploca chileae

1990 ◽  
Vol 36 (6) ◽  
pp. 438-448 ◽  
Author(s):  
Siegfried Maier ◽  
Horst Völker ◽  
Marita Beese ◽  
Victor A. Gallardo
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Fine Structure ◽  
Coastal Upwelling ◽  
The Other ◽  
Surface Topology ◽  
Cytoplasmic Material ◽  
Transmission Electron ◽  
Soft Bottoms ◽  
Sulfur Inclusions

Thioploca araucae and Thioploca chileae from the sublittoral soft bottoms of the coastal upwelling ecosystem off central Chile were examined by scanning and transmission electron microscopy. Except for filament diameter (30–43 and 12–20 μm, respectively) and slight differences in other dimensions and surface topology, the details of fine structure were essentially identical in the two species. The wall consisted of five layers, and only the inner layer was present in the septum. Multiple membrane intrusions dissected the procaryotic cytoplasmic material, which was restricted to a relatively thin layer within the wall. Sulfur inclusions and two other extracytoplasmic inclusions, as well as one kind of intracytoplasmic inclusion, were described. The central part of each cell consisted of one large vacuole, extending from septum to septum and representing a volume at least more than three times larger than the combined volume of wall and cytoplasm. The vacuole was separated from the other cell parts by a membrane. Electron-dense material was deposited between vacuole membrane and plasma membrane, between plasma membrane and wall, and inside membrane intrusions. A continuity between the vacuolar membrane and the other membranes was never encountered. The possible origin of such an extracytoplasmic membrane is discussed. Key words: benthos, electron microscopy, fine structure, Thioploca.

Ultrastructural assessment of cryofractured primate spermatozoon by analytical scanning electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 246-247
Author(s):  
R. P. Apkarian ◽  
K. Gopalkrishnan
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Single Sample ◽  
High Magnification ◽  
Transmission Electron ◽  
Large Numbers ◽  
Imaging Mode ◽  
Analytical Scanning Electron Microscopy ◽  
Scanning Electron

A cryofracture technique for scanning electron microscopy (SEM) was developed to provide an ultrastructural assessment of large numbers of chimpanzee sperm from a single centrifuged pellet. Normal spermatozoon observed in an analytical SEM at high magnification were used to determine the suitability of this technique for preserving the ultrastructural features of many sperm fractured through different regions. Although transmission electron microscopy (TEM) provides fine ultrastructural imaging of primate spermatozoon, this imaging mode does not provide large numbers of sperm in a single sample which can be rapidly scanned for profiles of ultrastructural abnormalities useful in infertility screening.

scholarly journals The localization of chitin synthase mediates the patterned deposition of chitin in developing Drosophila bristles

2019 ◽  
Author(s):  
Paul N. Adler
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Complex Structure ◽  
Chitin Synthase ◽  
Cell Type ◽  
Body Regions ◽  
Transmission Electron ◽  
Successful Group ◽  
Sensory Bristles ◽  
Parallel Arrays

AbstractThe insect exoskeleton is a complex structure that is a key for the life style of this very successful group of animals. It contains proteins, lipids and the N-acetyl glucosamine polymer chitin. Chitin is synthesized by the enzyme chitin synthase. In most body regions, chitin fibrils are found in a stack of parallel arrays that can be detected by transmission electron microscopy. Each array is rotated with respect to the layers above and below. In sensory bristles, chitin primarily accumulates in bands parallel to the proximal/distal axis of the bristle. These bands are visible by confocal microscopy providing experimental advantages. We have used this cell type and an edited chitin synthase gene to establish that the bands of chitin are closely associated with stripes of chitin synthase, arguing the localization of chitin synthase plays an important role in mediating the patterned deposition of chitin. This is reminiscent of what has been seen for chitin and chitin synthase in fungi and between cellulose and cellulose synthase in plants. Several genes are known to be essential for proper chitin deposition. We found one of these, Rab11 is required for the insertion of chitin synthase into the plasma membrane and a second, duskylike is required for plasma membrane chitin synthase to localize properly into stripes. We also established that the actin cytoskeleton is required for the proper localization of chitin synthase and chitin in developing sensory bristles.

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