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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 136 (1973), S. 433-446 
    ISSN: 1432-0878
    Schlagwort(e): Amoeba proteus ; Cytotic membrane turnover ; Permanent and induced endocytosis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Beschreibung / Inhaltsverzeichnis: Zusammenfassung Zur Bestimmung des cytotischen Membran-Turnovers wurden morphometrische Messungen an über 60 Zellen der Art Amoeba proteus durchgeführt. Danach nehmen diese Amöben 0,14% ihrer Zelloberfläche pro Minute durch permanente Endocytose in das Cytoplasma auf. A. proteus benötigt also insgesamt 12 Std, um die gesamte Zellmembran während der normalen Bewegung einmal zu erneuern. Infolge des geringen Membranturnovers kann der permanenten Endocytose keine aktive Bedeutung für die Erzeugung der Bewegungstriebkraft zugesprochen werden. In Übereinstimmung mit dieser Vermutung ließ sich eine Abhängigkeit zwischen Fortbewegungsgeschwindigkeit und Endocytoseintensität nicht nachweisen. Entsprechende Messungen mit drei verschiedenen Endocytoseinduktoren ergaben für die induzierte Endocytose in Abhängigkeit von der verwendeten Substanz eine wesentlich höhere Ingestionsrate von 0,43–2,25%/min. Derartige Spitzenwerte können allerdings nur innerhalb eng begrenzter Zeiträume von 15–30 min erzielt werden. Vergleicht man dagegen die Membranaufnahme während der permanenten und induzierten Endocytose über längere Zeitintervalle (4–5 Std), so bleibt die induzierte Endocytose mit 0,05–0,12%/min in der Intensität deutlich hinter der permanenten Endocytose (0,14%/min) zurück. Eine Erhöhung der Temperatur auf 30° und eine Erniedrigung auf 15°C bringen beide Endocytoseformen zum Erliegen. Die permanente Endocytose muß bei Amöben neben der Phagocytose als der wichtigste Mechanismus zur kontinuierlichen Aufnahme gelöster und suspendierter Stoffe (bis zur Größenordnung von Bakterien) angesehen werden.
    Notizen: Summary Cytotic membrane turnover of Amoeba proteus was morphometrically studied in more than 60 cells. The results obtained indicate that 0.14% of the total cell membrane area per minute is ingested by permanent endocytosis. Consequently during normal locomotion the total cell membrane area is renewed once within 12 hours. This rate is too low to play any role in the generation of motive force. No correlations were found between the rates of locomotion and permanent endocytosis. Comparative measurements on cells treated with three different substances inducing endocytosis reveal that induced endocytosis leads to an increased rate of membrane ingestion of 0.43–2.25%/min depending on the substance used. These high rates, however, are only maintained during short periods of time (15–30 min). When the rates are calculated on the basis of long periods of time (4–5 hours), it is obvious that induced endocytosis (0.05–0.12%/min) is less effective in long term membrane turnover than permanent endocytosis (0.14%/min). Endocytotic activity is completely abolished by both the increase and decrease in temperature to 30°C and 15°C respectively. In addition to discontinuous phagocytosis permanent endocytosis is an important mechanism for continuous ingestion of fluid including particles up to the size of bacteria.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 137 (1973), S. 97-109 
    ISSN: 1432-0878
    Schlagwort(e): Amoeba proteus ; Adsorptive function of mucoid layer ; Significance for induced endocytosis ; Electron microscopy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Beschreibung / Inhaltsverzeichnis: Zusammenfassung Die Mucoidschicht von Amoeba proteus enthält saure Mucopolysaccharide und ist funktionell einem Kationenaustauscher vergleichbar. Schwermetallpartikel und Proteine werden in Abhängigkeit vom pH-Wert des Kulturmediums in unterschiedlich starken Mengen an die Mucoidfilamente gebunden. Die dem Plasmalemm unmittelbar aufliegende globuläre Grundschicht besitzt dagegen für die untersuchten Substanzen keine Adsorptionsfunktion und unterscheidet sich demnach sowohl physiologisch als auch chemisch von der filamentartigen Zone. Quantitative Messungen über die pH-abhängige Anlagerung von Ferritin haben ergeben, daß die Mucoidfilamente in der Lage sind, Kationen aus dem Kulturmedium bis zu einer um das 17fache höheren Konzentration anzureichern. Die Ergebnisse der vorliegenden Untersuchung sprechen dafür, daß die Anreicherung derartiger Substanzen an der Zelloberfläche eine notwendige und physiologisch sinnvolle Voraussetzung für die Auslösung einer induzierten Endocytose darstellt.
    Notizen: Summary The mucoid layer of Amoeba proteus contains acid mucopolysaccharides, which are involved in the exchange of cations. Depending on the pH of the external medium, different amounts of heavy metal particles and proteins are bound to the mucoid filaments. The globular ground layer, which is directly apposed to the plasma membrane does not bind any of the substances studied and, therefore, differs from the mucoid filaments in function as well as chemical nature. Measurements of the pH-dependent adsorption of ferritin demonstrate that the mucoid filaments are able to accumulate cations in concentrations 17 times that of the external medium. These results suggest that the accumulation of substances at the cell surface is a prerequisite for the initiation of induced endocytosis.
    Materialart: Digitale Medien
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  • 3
    ISSN: 1615-6102
    Schlagwort(e): Amoeba proteus ; Cytoskeleton ; Morphological changes ; Iodoacetamido-fluorescein-(IAF)-labeled actin ; Microinjection
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Iodoacetamido-fluorescein-(IAF)-labeled actin was microinjected into normal locomotingAmoeba proteus. Thereafter (30–60 minutes) changes in the cytoplasmic fluorescence distribution pattern and contractile activity were induced by internal and external chemical stimulation. Different agents such as phalloidin, procaine, 2.4-dinitrophenol (DNP), puromycin, ouabain and n-ethyl maleimide (NEM) interfere with the excitation-contraction mechanism involved in ordered pseudopodium formation during ameboid movement and cause various morphogenetic reactions based on actin polymerization-depolymerization cycles. Most frequent changes are (a) local condensation of IAF-actin and formation of a continuous IAF-actin layer at the cytoplasmic surface of the cell membrane and around the pulsating vacuole, (b) immobilization and hyalo-granuloplasm separation by combined contraction and detachment of the IAF-actin layer from the cell membrane, (c) organized and disorganized formation of pseudopodia by local contraction and disintegration of the IAF-actin layer, and (d) alterations in the rheological properties of the protoplasmic matrix by changes in the molecular state of soluble actin not incorporated into the cytoskeleton. The experimental approaches to the function of the actomyosin system in large amebas attainable by the method ofin vivo molecular cytochemistry are discussed in detail with respect to the participation of the cytoskeleton in motive force generation for cytoplasmic streaming and ameboid movement.
    Materialart: Digitale Medien
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  • 4
    ISSN: 1615-6102
    Schlagwort(e): Mucous layer ; Receptor sites ; Cations ; Lectins ; Amoeba proteus ; Fluorescent analog cytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Two different receptor sites, located on the cell surface ofAmoeba proteus were detected by using fluorescent analog cytochemistry (FAC) and electron microscopy (EM). Bovine serum albumin labeled with fluoresceine-isothiocyanate (FITC-BSA) and unlabeled ferritin bind, in a pH-dependent manner, as cations at the outer filaments of the mucous layer. The anionic receptor sites show a high affinity for Ca-ions which suppress the binding capacity of FITC-BSA and ferritin at low pH-values. The cation receptors obviously play an important role in the initiation of pinocytosis as demonstrated by the internalization, intracellular translocation and sequestration of the FITC-BSA. FITC- or ferritin-labeled concanavalin A (FITC-Con A, ferritin-Con A) bind predominantly in a pH-independent manner at the tips of the outer filaments and the basal zone of the mucous layer. The binding capacity of FITC-Con A is not influenced by external Ca-ions. Other lectins such asDolichos bifloris agglutinin (DBA), peanut agglutinin (PNA),Ricinus communis agglutinin I (RCA I), soybean agglutinin (SBA),Ulex europaeus agglutinin I (UEA I) and wheat germ agglutinin (WGA) are not specifically bound to the cell surface. So far, no experimental evidence has been gathered for the definitive function of a Con-A receptor in the mucos layer ofAmoeba proteus.
    Materialart: Digitale Medien
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  • 5
    Digitale Medien
    Digitale Medien
    Springer
    Protoplasma 147 (1988), S. 117-124 
    ISSN: 1615-6102
    Schlagwort(e): Heat-shock ; Contractile layers ; Intracellular movements ; Amoeba proteus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Ultrastructural and immunocytochemical studies allow the localization and identification of a microfilament cortex in heat-shockedAmoeba proteus at different stages of recovery to room temperature. Immediately after heating the cortex is in close contact with the cytoplasmic face of the plasma membrane; however, during cooling it detaches from the membrane and shifts toward the cell centre thus separating a region of peripheral hyaloplasm from central granuloplasm. After polymerization of a new submembrane cortex several detachment and reformation cycles rhythmically repeated for 2–3 hours until a multitude of stratified layers has been formed in the hyaloplasm. Electron micrographs reveal that the cortical layer at the plasma membrane is merely composed of a network of actin filaments, whereas the retracted contractile layers in the hyaloplasm and at the granuloplasmic border contain both, thick and thin filaments often arranged in bundles. The heat-shock induced activities of the microfilament cortex are based on the highly contractile properties of this system in conjunction with controlled displacements in the equilibrium between F- and G-actin.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 6
    ISSN: 1615-6102
    Schlagwort(e): Cytoskeleton ; Cytoplasmic organization ; Cell organelle transport ; Fluorescence microscopy ; Spongilla lacustris
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The cytoskeleton in epithelial cells ofSpongilla lacustris is constructed of microtubules radiating from the nuclear region and terminating at the cell periphery as well as microfilaments forming a cortical layer beneath the plasma membrane and distinct fibers in the cytoplasmic matrix. Single frame analysis and in vivo labeling of mitochondria with Rh 123, endosomes or lysosomes with TRITC-BSA, endoplasmic reticulum (ER) with DiOC6 (3) and dictyosomes with C6-NBD-ceramide points to the microtubular system as a candidate for controlled cytoplasmic organization and active transport of these cell organelles. In epithelial cells with an intact microtubular system, mitochondria and endosomes or lysosomes show a regular shuttle movement between the nucleus and the cell periphery with a velocity of 1.3–1.4 μm/s; the ER forms a more or less dynamic two-dimensional network in the entire cytoplasmic matrix, and dictyosomes are arranged in a ring-like manner around the nucleus. In epithelial cells treated with colchicine or colcemid, mitochondria and endosomes or lysosomes gather in the perinuclear region and become immobile; the ER accumulates near the cell center, whereas most dictyosomes distribute randomly over the whole cytoplasm. Transformation of the microfilament system with cytochalasin D has no influence on cell organelle distribution and dynamics but impedes cell locomotion and cell surface activities.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 7
    ISSN: 1615-6102
    Schlagwort(e): Lipid pathways ; Fluorescent analogues ; Lysosomes ; Golgi apparatus ; Amoeba proteus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The uptake and intracellular transport of 5 different lipid analogues derived from phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, ceramide, and cholesterol have been studied in livingAmoeba proteus using fluorescence microscopy. Phosphatidylcholine and sphingomyelin are predominantly transported from the external environment to the cell interior in a manner consistent with induced macropinocytosis. On the other hand, phosphatidylethanolamine, ceramide, and cholesterol mainly enter the cellular matrix by carrier-mediated, ATP-dependent transmembrane transport. In general, all lipid analogues are first imported to a large pool of endosomal or lysosomal vacuoles, and then partitioned to numerous tiny cisternal elements; only sphingomyelin remains in the lysosomes and is not exported to other membrane compartments. The ultrastructural localization of ceramide indicates that the cisternal elements result from the decay of the Golgi apparatus into single cisternae during lipid accumulation. As a whole, the transport pathway of lipid analogues inA. proteus from the cell surface to different cell organelles shows many similarities to respective processes in a variety of metazoan cells.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 8
    Digitale Medien
    Digitale Medien
    Springer
    Protoplasma 100 (1979), S. 33-43 
    ISSN: 1615-6102
    Schlagwort(e): Amoeba proteus ; Ca++-binding sites ; Cytochemical demonstration ; Induced pinocytosis ; Plasma membrane
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Different methods were used to demonstrate the existence of Ca++-binding sites (Ca++-bs) at the plasma membrane ofAmoeba proteus. In pinocytoting animals the number (indicated by the average distanced in nm) and size (average longitudinal axiss in nm) of Ca++-bs at the cytoplasmic surface of the cell membrane were significantly increased (d=162±15;n=41 ands=93±5;n=47) in comparison to controls (d=208 ±21;n=37 ands=59±8;n=45). The ratio of P: Ca obtained by X-ray microanalysis was in the range of 1.5. The differences observed in the two experimental groups of amoebae are explained by conformational changes in the molecular structure and an increased Ca++-permeability of the plasma membrane during induced pinocytosis. Microplasmodia of the acellular slime moldPhysarum polycephalum investigated for comparison were found to have no Ca++-bs at the interior cell surface.
    Materialart: Digitale Medien
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  • 9
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 197 (1979), S. 263-279 
    ISSN: 1432-0878
    Schlagwort(e): Induced pinocytosis ; Dynamics ; Motive force generation ; Light and electron microscopy ; Amoeba proteus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The mechanism of induced pinocytosis was investigated in Amoeba proteus by light and electron microscopy. The application of nine different inducing substances revealed that pinocytotic channel formation, elongation, vesiculation, shortening and disappearance are the result of the successive or simultaneous action of both traction and pressure forces, which are produced by the contractile activity of a plasma membrane-associated layer of filaments ranging from a few hundred nm to several μ in thickness. The initial phase of channel formation is caused by traction forces according to the membrane flow concept, whereas channel elongation and vesiculation mainly result from pressure forces in conjunction with the extrusion of small hyaline pseudopodia. Shortening and disappearance of the pinocytotic channels are brought about by local contractions of the cortical filament layer in the basal region of the hyaline pseudopodia. Experiments using latex beads as marker particles together with inducing substances show that a rapid membrane turnover during pinocytosis can be excluded, and that the plasma membrane slides as an entire structure over the underlying cytoplasm.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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