Membrane recycling at the cytoproct of Tetrahymena

1979 ◽  
Vol 35 (1) ◽  
pp. 217-227
Author(s):  
R.D. Allen ◽  
R.W. Wolf
Keyword(s):  
Plasma Membrane ◽  
Single Cells ◽  
Tetrahymena Pyriformis ◽  
Food Vacuole ◽  
Membrane Recycling ◽  
Vacuole Membrane ◽  
Thin Section Electron Microscopy ◽  
Section Electron ◽  
Food Vacuoles ◽  
Spherical Vesicles

Exocytosis and membrane recycling at the cytoproct (cell anus) of Tetrahymena pyriformis were studied using thin-section electron microscopy. Single cells were fixed at specific times relative to the elimination of the vacuole's contents—before elimination, at elimination, 3–5 s and 10–15 s following elimination. The closed cytoproct is distinguished from other pellicular regions by a single membrane at the cell surface which is circumscribed by an electron-opaque flange that links or welds the plasma membrane to the underlying alveolar margins. Microtubules originating in the flange pass inward where they lie over, and possibly guide, the approaching food vacuoles to the cytoproct. Food facuoles near the cytoproct are also accompanied by coats of microfilaments. These microfilaments appear to be active in the channelling and endocytosis of food vacuole membrane. Upon cytoproct opening the plasma membrane and food vacuole membrane fuse. Elimination seems to be essentially passive and is accomplished by re-engulfment of the old food vacuole membrane which is constantly associated with microfilaments. Reengulfment of all the food vacuole membrane requires 10–15 s and results in a closed cytoproct. The membrane remnants embedded in microfilaments form a cluster under the closed cytoproct. At the periphery of this cluster remnants take the shape of 70–130-nm spherical vesicles or 0.2-micrometer-long flattened vesicles.

scholarly journals The Cytoproct of Paramecium Caudatum: Structure and Function, Microtubules, and Fate of Food Vacuole Membranes

1974 ◽  
Vol 14 (3) ◽  
pp. 611-631
Author(s):  
RICHARD D. ALLEN ◽  
R. W. WOLF
Keyword(s):  
Plasma Membrane ◽  
Single Cells ◽  
Membrane Vesicles ◽  
Food Vacuole ◽  
Paramecium Caudatum ◽  
Vacuole Membrane ◽  
Narrow Ridge ◽  
Membrane Changes ◽  
And Function ◽  
Over The Top

The cytoproct or cell anus of Paramecium caudatum was studied, using light optics and electron microscopy, at known times before, during and following food vacuole egestion. This was accomplished by microscopically observing single cells, fixing these cells at specific times and finally serial sectioning these individually processed cells. The cytoproct, at rest, is a long narrow ridge along the posterior suture. It contains 2 uniquely positioned components which identify this structure as the cytoproct: piles of fibres along the inside surfaces of the ridge, and microtubules passing from the epiplasm at the summit of the ridge down into the cytoplasm. The plasma membrane is continuous over the top of the ridge. The cortical basal bodies adjacent to the ridge have bundles of microtubules passing into the cytoplasm from an opaque plaque at their proximal ends. These 2 sets of microtubules may function in guiding the food vacuoles to the cytoproct. A model is presented in which motive forces generated between the microtubules and the food vacuole membrane bring the food vacuole to the cytoproct and, in addition, pull the cytoproct lips apart so that the food vacuole membrane and plasma membrane come into contact and fuse together, thus opening the food vacuole to the outside. The plasma membrane increases in area between the parting lips, possibly, as the result of intercalation of membrane vesicles into the plasma membrane at the top of the ridge. Immediately after this opening is formed the food vacuole membrane changes from a smooth topography to a highly convoluted one. The membrane is engulfed through a process of endocytosis resulting in an accumulation of membranous fragments in the cytoplasm below the cytoproct. The endocytic forces probably bring about the restitution of the cytoproct ridge by pulling the lips back together as the membrane is engulfed. A filamentous meshwork underlying the food vacuole membrane may be active in this endocytic process.

Deformation of Yeast Plasma Membrane by Ethidium Bromide

1988 ◽  
Vol 46 ◽  
pp. 254-255
Author(s):  
E. Keyhani
Keyword(s):  
Plasma Membrane ◽  
Thin Section ◽  
Ethidium Bromide ◽  
Freeze Fracture ◽  
Mutagenic Effect ◽  
Yeast Cells ◽  
Hydrophobic Region ◽  
Thin Section Electron Microscopy ◽  
Section Electron ◽  
Deep Pits

The mutagenic effect of ethidium bromide on the mitochondrial DNA is well established. Using thin section electron microscopy, it was shown that when yeast cells were grown in the presence of ethidium bromide, besides alterations in the mitochondria, the plasma membrane also showed alterations consisting of 75 to 110 nm-deep pits. Furthermore, ethidium bromide induced an increase in the length and number of endoplasmic reticulum and in the number of intracytoplasmic vesicles.Freeze-fracture, by splitting the hydrophobic region of the membrane, allows the visualization of the surface view of the membrane, and consequently, any alteration induced by ethidium bromide on the membrane can be better examined by this method than by the thin section method.Yeast cells, Candida utilis. were grown in the presence of 35 μM ethidium bromide. Cells were harvested and freeze-fractured according to the procedure previously described.

scholarly journals STUDIES OF THE ULTRASTRUCTURE AND RIBOSOMAL ARRANGEMENTS OF THE PLEUROPNEUMONIA-LIKE ORGANISM A5969

1965 ◽  
Vol 25 (1) ◽  
pp. 139-150 ◽  
Author(s):  
Jack Maniloff ◽  
Harold J. Morowitz ◽  
Russell J. Barrnett
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Chemical Analysis ◽  
Thin Section ◽  
Differential Centrifugation ◽  
Thin Section Electron Microscopy ◽  
Section Electron ◽  
Section Electron Microscopy ◽  
Cellular Organelles

Thin-section electron microscopy, together with isolation of cellular organelles by differential centrifugation and chemical analysis, has been used to investigate the ultrastructure of the avian pleuropneumonia-like organism A5969. Each cell (approximate diameter 5500 A) was surrounded by a 150 A plasma membrane. In the center of the cell was an unbounded area, granular in appearance and containing the cell's DNA. The periphery of the cell contained granules of several different sizes and densities. The most dense particles (150 A) corresponded to the 78S ribosomes. These particles exhibited two predominant arrangements: (a) sometimes they showed cubic packing; (b) most arrays, however, were consistent with cylindrical arrangements of approximately 50 particles. Bundles of up to 18 arrays were observed. Structured blebs have been found protruding from the surface of log phase cells.

scholarly journals Phagotrophy and Two New Structures in the Malaria Parasite Plasmodium berghei

1959 ◽  
Vol 6 (1) ◽  
pp. 103-112 ◽  
Author(s):  
Maria A. Rudzinska ◽  
William Trager
Keyword(s):  
Plasma Membrane ◽  
Plasmodium Berghei ◽  
Food Vacuole ◽  
The Body ◽  
Host Cells ◽  
Butyl Methacrylate ◽  
Host Cytoplasm ◽  
Mitochondrial Functions ◽  
Food Vacuoles ◽  
Almost All

Blood collected from rats infected with Plasmodium berghei was centrifuged and the pellet was fixed for 1 hour in 1 per cent buffered OsO4 with 4.9 per cent sucrose. The material was embedded in n-butyl methacrylate and the resulting blocks sectioned for electron microscopy. The parasites were found to contain, in almost all sections, oval bodies of the same density and structure as the host cytoplasm. Continuity between these bodies and the host cytoplasm was found in a number of electron micrographs, showing that the bodies are formed by invagination of the double plasma membrane of the parasite. In this way the host cell is incorporated by phagotrophy into food vacuoles within the parasite. Hematin, the residue of hemoglobin digestion, was never observed inside the food vacuole but in small vesicles lying around it and sometimes connected with it. The vesicles are pinched off from the food vacuole proper and are the site of hemoglobin digestion. The active double limiting membrane is responsible not only for the formation of food vacuoles but also for the presence of two new structures. One is composed of two to six concentric double wavy membranes originating from the plasma membrane. Since no typical mitochondria were found in P. berghei, it is assumed that the concentric structure performs mitochondrial functions. The other structure appears as a sausage-shaped vacuole surrounded by two membranes of the same thickness, density, and spacing as the limiting membrane of the body. The cytoplasm of the parasite is rich in vesicles of endoplasmic reticulum and Palade's small particles. Its nucleus is of low density and encased in a double membrane. The host cells (reticulocytes) have mitochondria with numerous cristae mitochondriales. In many infected and intact reticulocytes ferritin was found in vacuoles, mitochondria, canaliculi, or scattered in the cytoplasm.

scholarly journals FOOD VACUOLE MEMBRANE GROWTH WITH MICROTUBULE-ASSOCIATED MEMBRANE TRANSPORT IN PARAMECIUM

1974 ◽  
Vol 63 (3) ◽  
pp. 904-922 ◽  
Author(s):  
Richard D. Allen
Keyword(s):  
Membrane Surface ◽  
Three Dimensional ◽  
Food Vacuole ◽  
Structural System ◽  
Digestive Vacuole ◽  
Membrane Surface Area ◽  
Vacuole Membrane ◽  
Membrane Growth ◽  
Food Vacuoles ◽  
Membrane Disk

Evidence from a morphological study of the oral apparatus of Paramecium caudatum using electron microscope techniques have shown the existence of an elaborate structural system which is apparently designed to recycle digestive-vacuole membrane. Disk-shaped vesicles are filtered out of the cytoplasm by a group of microtubular ribbons. The vesicles, after being transported to the cytostome-cytopharynx region in association with these ribbons, accumulate next to the cytopharynx before they become fused with the cytopharyngeal membrane. This fusion allows the nascent food vacuole to grow and increase its membrane surface area. The morphology of this cytostome-cytopharynx region is described in detail and illustrated with a three-dimensional drawing of a portion of this region and a clay sculpture of the oral apparatus of Paramecium. Evidence from the literature for the transformation of food vacuole membrane into disk-shaped vesicles both from condensing food vacuoles in the endoplasm and from egested food vacuoles at the cytoproct is presented. This transformation would complete a system of digestive vacuole membrane recycling.

Dual capacity for nutrient uptake in Tetrahymena. V. Utilization of amino acids and proteins

1979 ◽  
Vol 36 (1) ◽  
pp. 343-353
Author(s):  
E. Orias ◽  
L. Rasmussen
Keyword(s):  
Amino Acids ◽  
Plasma Membrane ◽  
Tetrahymena Thermophila ◽  
Amino Acid Uptake ◽  
Food Vacuole ◽  
Transport Systems ◽  
Acid Uptake ◽  
Vacuole Formation ◽  
Carrier Mediated Transport ◽  
Food Vacuoles

We investigated the relative contributions of phagocytosis and plasma membrane transport to the uptake of amino acids and a protein (egg albumin) in amounts which allow Tetrahymena thermophila to grow and multiply. We used a mutant capable of indefinite growth without food vacuole formation (phagocytosis) and its wild type (phagocytosis-competent) isogenic parental strain. Our results suggest that phagocytosis is not required for free amino acid uptake, most or all of which can be attributed to carrier-mediated transport systems, apparently located on the plasma membrane. In contrast, phagocytosis is required for utilization of the protein. Proteins can supply required amino acids in amounts sufficient for growth only when food vacuoles are formed. We conclude that Tetrahymena thermophila either possesses no endocytic mechanisms at the cell surface other than food vacuole formation or, if it does, these putative mechanisms are not capable of nutritionally meaningful rates of protein uptake.

Exocytosis, endocytosis and membrane recycling in Tetrahymena thermophila

1988 ◽  
Vol 89 (4) ◽  
pp. 515-520
Author(s):  
ARNO TIEDTKE ◽  
PETER HÜNSELER ◽  
JORGE FLORIN-CHRISTENSEN ◽  
MONICA FLORIN-CHRISTENSEN
Keyword(s):  
Cell Lines ◽  
Tetrahymena Thermophila ◽  
Food Vacuole ◽  
Secondary Effect ◽  
Membrane Recycling ◽  
Acid Hydrolase ◽  
Wild Type ◽  
Acid Hydrolases ◽  
Vacuole Formation ◽  
Food Vacuoles

Mutant and wild-type cell lines of Tetrahymena thermophila were used to investigate a possible connection between acid hydrolase secretion and the major processes through which membranes are recycled in this ciliated protozoon. These processes consist of food vacuole formation (endocytosis), and food vacuole egestion and mucocyst release (both exocytosis). We have found that a mutant (MS-1, see−) blocked in hydrolase secretion is not blocked in either food vacuole formation or egestion and that it has normal mucocyst exocytosis. Another line of experiments with wild-type cells showed also that hydrolase secretion and endocytosis are independent of each other. Thus, sucrose (0.1m) did not interfere with hydrolase secretion, but blocked food vacuole formation. Furthermore, release of acid hydrolases was selectively stimulated by dibucaine without any effect on food vacuole egestion. Finally, exocytosis of mucocysts could occur without simultaneous release of acid hydrolases, as when cells were exposed to (0.15M-NaCl, which evokes a massive secretory response of mucocysts. Our results demonstrate that formation and egestion of food vacuoles and exocytosis of mucocysts are unrelated to secretion of acid hydrolases. Furthermore, they suggest that secretion of acid hydrolases is not a secondary effect of membrane recycling through these processes.

Feeding in Ciliated Protozoa

1974 ◽  
Vol 15 (2) ◽  
pp. 379-401
Author(s):  
JOHN. A. KLOETZEL
Keyword(s):  
Cell Surface ◽  
Buccal Cavity ◽  
Surface Membrane ◽  
Food Vacuole ◽  
Food Ingestion ◽  
Membrane Flow ◽  
Vacuole Membrane ◽  
Vacuole Formation ◽  
Food Vacuoles ◽  
Lamellar Material

The ciliate Euplotes is able to expend a very large amount of membrane in the formation of food vacuoles. Calculations based on the rate of ingestion of the food organism Tetrahymena indicate that an amount of food vacuole membrane equivalent to approximately 50-150% of the total Euplotes cell surface area can be produced within 5-10 min. An aggregation of osmiophilic, membrane-limited ‘pharyngeal disks’ is found packed in the cytoplasm just beneath the cell surface membrane in the region of the cell mouth and cytopharynx. These disks, which can be seen also in living cells, have average dimensions of 2 µm diameter by 100 nm thickness, and contain tightly packed layers of a thin lamellar material. Electron micrographs have revealed the apparent fusion of the limiting membrane of disks with the cell's plasma membrane at the base of the gullet. The lamellar disk contents are thereby released to the exterior medium in the buccal cavity, where they form a loosely packed layer over the surface membrane. It is postulated that the pharyngeal disks represent a repository of preformed membrane for use in food vacuole formation. The disk contents may also play a role in food ingestion, although this is not well defined at present. The myeloid content of old food vacuoles is very similar to that of nearby disks in the cytoplasm, suggesting that the disks may form by pinching from shrinking food vacuoles during the digestive cycle. Thus a cycle of membrane flow is envisaged, with the pharyngeal disks (1) coalescing with the surface membrane during food vacuole formation, (2) reforming by pinching from these food vacuoles during digestion, and (3) migrating back to the oral region to serve as a membrane store for subsequent food vacuole formation.

Cell Multiplication in Tetrahymena Cultures after Addition of Particulate Material

1973 ◽  
Vol 12 (1) ◽  
pp. 275-286
Author(s):  
L. RASMUSSEN ◽  
L. MODEWEG-HANSEN
Keyword(s):  
Tetrahymena Pyriformis ◽  
Food Vacuole ◽  
Particulate Material ◽  
Cell Multiplication ◽  
Particulate Suspensions ◽  
Proteose Peptone ◽  
Generation Times ◽  
Food Vacuoles ◽  
Net Charges ◽  
Peptone Broth

We have studied the effects of adding particulate supplements to populations of Tetrahymena pyriformis in 2% sterile-filtered proteose peptone broth which supports cell multiplication poorly (generation times in excess of 40 h). The tested compounds were: heat-sterilized suspensions of egg albumin, nutritionally inert particles of polystyrene, sulphopropyl and quarternary amino-ethyl substituted dextran (in concentrations of 4, 40 and 400 µg per ml). The particles had approximately the same size, but differed in their electric net charges. Particulate suspensions of 40 µg per ml or more greatly improved cell multiplication rates (generation times about 6 h). It is probable that the effect of the particles is to induce formation of food vacuoles without which cell multiplication and growth is very slow. The contribution of the food vacuole to nutrient uptake in Tetrahymena is discussed.

scholarly journals ASSEMBLY OF LIPIDS INTO MEMBRANES IN ACANTHAMOEBA PALESTINENSIS

1971 ◽  
Vol 50 (3) ◽  
pp. 634-651 ◽  
Author(s):  
Francis J. Chlapowski ◽  
R. Neal Band
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Specific Activity ◽  
Food Vacuole ◽  
Cell Fractionation ◽  
Membrane Phospholipids ◽  
Golgi Membranes ◽  
Cytoplasmic Vesicles ◽  
Food Vacuoles ◽  
Phospholipid Pool

The membranes of Acanthamoeba palestinensis were studied by examination in fixed cells, and then by following the movements of glycerol-3H-labeled phospholipids by cell fractionation. Two previously undescribed structures were observed: collapsed cytoplasmic vesicles of cup shape, and plaques in food vacuole and plasma membrane similar in size to the collapsed vesicles. It appeared that the plaques formed by insertion of collapsed vesicles into membranes and/or that collapsed vesicles formed by pinching off of plaques. Fractions were isolated, enriched with nuclei, rough endoplasmic reticulum (RER), plasma membrane, Golgi-like membranes, and collapsed vesicles. The changes in specific activity of glycerol-3H-labeled phospholipids in these membranes during incorporation, turnover, and after pulse-labeling indicated an ordered sequence of appearances of newly synthesized phospholipids, first in nuclei and RER, then successively in Golgi membranes, collapsed vesicles, and finally, plasma membrane. In previous work we had found no large nonmembranous phospholipid pool in A. palestinensis. These observations are consistent with the hypothesis that membrane phospholipids are synthesized, perhaps as integral parts of membranes, in RER and nuclei. Subsequently, some of the newly synthesized phospholipids are transported to the Golgi complex to become integrated into the membranes of collapsed vesicles, which are precursors of the plasma membrane. Collapsed vesicles from the plasma membrane by inserting into it as plaques. When portions of the plasmalemma from food vacuoles, collapsed vesicles pinch off from their membranes and are recycled back to the cell surface.

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