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At least 127 records · Page 7Linked to original sources
PubMed1995

Dictyostelium myosin I double mutants exhibit conditional defects in pinocytosis.

The functional relationship between three Dictyostelium myosin Is, myoA, myoB, and myoC, has been examined through the creation of double mutants. Two double mutants, myoA-/B- and myoB-/C-, exhibit similar conditional defects in fluid-phase pinocytosis. Double mutants grown in suspension culture are significantly impaired in their ability to take in nutrients from the medium, whereas they are almost indistinguishable from wild-type and single mutant strains when grown on a surface. The double mutants are also found to internalize gp126, a 116-kD membrane protein, at a slower rate than either the wild-type or single mutant cells. Ultrastructural analysis reveals that both double mutants possess numerous small vesicles, in contrast to the wild-type or myosin I single mutants that exhibit several large, clear vacuoles. The alterations in fluid and membrane internalization in the suspension-grown double mutants, coupled with the altered vesicular profile, suggest that these cells may be compromised during the early stages of pinocytosis, a process that has been proposed to occur via actin-based cytoskeletal rearrangements. Scanning electron microscopy and rhodamine-phalloidin staining indicates that the myosin I double mutants appear to extend a larger number of actin-filled structures, such as filopodia and crowns, than wild-type cells. Rhodamine-phalloidin staining of the F-actin cytoskeleton of these suspension-grown cells also reveals that the double mutant cells are delayed in the rearrangement of cortical actin-rich structures upon adhesion to a substrate. We propose that myoA, myoB, and myoC play roles in controlling F-actin filled membrane projections that are required for pinosome internalization in suspension.

Actins↗
PubMed1975

Quantitative studies of pinocytosis. I. Kinetics of uptake of (125I)polyvinylpyrrolidone by rat yolk sac cultured in vitro.

A method is described for the in vitro culture of 17.5-day rat visceral yolk sac. Tissue survival was good as judged by light and electron microscopy. The rate of pinocytic uptake of 125I-labeled polyvinylpyrrolidone by the tissue was constant both within and between experiments. Within the concentration range 0.15-24 mug/ml, the 125I-labeled polyvinylpyrrolidone neither stimulated nor inhibited pinocytosis. The system offers many advantages in the quantitative study of the physical basis of pinocytosis.

Animals↗
PubMed1982

Inhibition of pinocytosis in rat embryo fibroblasts treated with monensin.

Rat embryo fibroblasts cultured in the presence of monensin exhibited an inhibited uptake of horseradish peroxidase. The inhibition was detected after 3 h, after which time the cells became increasingly vacuolated; the concentration of monensin required to inhibit pinocytosis (0.4 microM for half-maximum inhibition at 18 h) was similar to that found by others to inhibit secretion. Both the exchange of 5'-nucleotidase between the membranes of cytoplasmic organelles and the cell surface and the internalization of anti-5'-nucleotidase bound to the cell surface were inhibited by approximately 90% in monensin-treated cells. The effects of monensin were reversible: cells cultured first with monensin, and then in fresh medium, exhibited control levels of horseradish peroxidase uptake, exchange of 5'-nucleotidase, and internalization of anti-5'-nucleotidase bound to the cell surface. After monensin treatment, the median density of both galactosyl transferase and 5'-nucleotidase increased from 1.128 to 1.148, and the median density of both N-acetyl-beta-glucosaminidase and horseradish peroxidase taken up by endocytosis decreased from 1.194 to 1.160. The results indicate that monensin is a reversible inhibitor of pinocytosis and, presumably, therefore, of membrane recycling. They suggest that the inhibition of membrane recycling occurs at a step other than the fusion of pinocytic vesicles with lysosomes and is perhaps a consequence of an effect of the ionophore on the Golgi complex.

5'-Nucleotidase↗
PubMed1983

Pinocytosis and intracellular degradation of exogenous protein: modulation by amino acids.

Intracellular degradation of exogenous (serum) proteins provides a source of amino acids for cellular protein synthesis. Pinocytosis serves as the mechanism for delivering exogenous protein to the lysosomes, the major site of intracellular degradation of exogenous protein. To determine whether the availability of extracellular free amino acids altered pinocytic function, we incubated monolayers of pulmonary alveolar macrophages with the fluid-phase marker, [14C]sucrose, and we dissected the pinocytic process by kinetic analysis. Additionally, intracellular degradation of endogenous and exogenous protein was monitored by measuring phenylalanine released from the cell monolayers in the presence of cycloheximide. Results revealed that in response to a subphysiological level of essential amino acids or to amino acid deprivation, (a) the rate of fluid-phase pinocytosis increased in such a manner as to preferentially increase both delivery to and size of an intracellular compartment believed to be the lysosomes, (b) the degradation of exogenously supplied albumin increased, and (c) the fraction of phenylalanine derived from degradation of exogenous albumin and reutilized for de novo protein synthesis increased. Thus, modulation of the pinosome-lysosome pathway may represent a homeostatic mechanism sensitive to the availability of extracellular free amino acids.

Amino Acids↗
PubMed1983

Reversible pinocytosis in polymorphonuclear leukocytes.

Since pinocytosis has only been recently recognized in polymorphonuclear leukocytes (PMNs), little is known about the fate of pinosomes. Here we report that pinosomes can fuse with the cytoplasmic granules of PMNs. We also find that at least for a short period of time after formation, pinosomes can fuse with the plasma membrane and release their contents to the outside. We present a morphological description and biochemical data on the kinetic parameters of a steady state pool of reversible pinosomes in PMNs. In addition, we have developed conditions under which pinosomes continue to form and fuse with the plasma membrane but fail to fuse with the cytoplasmic granules, i.e., only "reversible" pinocytosis occurs. This inhibition of fusion with the granules is not due to an inability of the pinosomes to move from the surface since under these conditions pinosomes labeled with an electron-dense marker can be seen in the cell interior.

Animals↗
PubMed1966

The regulation of pinocytosis in mouse macrophages. I. Metabolic requirements as defined by the use of inhibitors.

A method is described to study the formation of pinocytic vesicles in cultivated mouse macrophages. Vesicles which arise in the peripheral cytoplasm and are in transit to the centrosphere region are enumerated by the phase-contrast microscopy of glutaraldehyde-fixed cells. Under these conditions there is a prompt reversible response of vesicle formation to calf serum factors in the external environment. The reduction of pinosome formation by a variety of metabolic inhibitors was then studied in a medium containing 50% newborn calf serum. Inhibitors of both glycolysis and respiration reduced vesicle formation to low levels. This influence was most striking with cyanide, antimycin A, and anaerobiosis. This indicates an important contribution of an intact respiratory pathway in pinocytosis. Both 2,4-dinitrophenol and oligomycin inhibited vesicle formation at low concentrations. These data suggest a central role of ATP as an energy source for vesicle formation. Inhibitors of protein synthesis, such as puromycin and p-fluorophenylalanine, produced a prompt reduction in vesicle formation. The action of p-fluorophenylalanine was effectively reversed with L-phenylalanine. Actinomycin D depressed pinocytosis to low levels at concentrations of 0.1 to 0.003 microg/ml. This effect was observed only after a 120 min lag phase. A 10 degrees C reduction in ambient temperature lowered vesicle counts to 30% of control preparations at 37 degrees C.

Animals↗
PubMed1982

Inhibition of rat yolk sac pinocytosis by cadmium and its reversal by zinc.

The effect of cadmium on the in vitro pinocytic rate of 10- and 11-day-old rat yolk sacs grown on rat serum in culture was studied using the synthetic substrate 125I-labeled polyvinylpyrrolidone. In medium containing concentrations of added cadmium similar to those achieved in serum by intraperitoneal dosing of rats, the rate of pinocytosis in vitro was decreased by up to 55%. When zinc was included in the medium as well, the effect of cadmium was markedly reduced, and the rate of pinocytosis was restored to about normal. The results of this in vitro study suggest that cadmium can limit and supply of nutrients to the rat embryo in a nonspecific manner, and that elevated levels of zinc can restore the ability of the yolk sac to accumulate substances from the surrounding medium. Whereas cadmium may act as a teratogen at a number of sites, the demonstrated effect of this metal on the yolk sac provides a new insight into what could be a primary mode of action of cadmium on fetal development.

Animals↗
PubMed1984

Sequential production of Birbeck granules through adsorptive pinocytosis.

A case of vitiligo with inflammatory raised borders was observed electron microscopically, resulting in an interesting view of the formation of Birbeck granules in Langerhans cells. Following the formation of larger coated vesicles, which perform adsorptive pinocytosis from the cell membrane, membrane invagination of the cell occurred shown as the tubular infolding, resulting in the observation of the characteristic Birbeck granule band pattern in its interior. This phenomenon supports the theory of Hashimoto and Tarnowski (1968) that Birbeck granules are formed from the infolding of the cell membrane. In addition, our study shows the involvement of adsorptive pinocytosis in the formation of the granules. It was suggested that when the coated Birbeck granule shifts into the cell, possibly its coat is detached and the vesicle portion forms the globule of the Birbeck granule.

Adult↗
PubMed1967

Plasma membrane: substructural changes correlated with electrical resistance and pinocytosis.

Inducers of pinocytosis in amoeba cause as much as a 50-fold decrease in the electrical resistance of the plasma membrane prior to the formation of the typical tunnels and vacuoles. In this state the thickness of the electron-transparent core or lamella of the unit membrane is at least twice as thick as that of the control. The changes in structure and resistance as well as the induction of pinocytosis are dependent on the initial external concentration of calcium. These changes are rapidly reversed when the concentration of calcium in the external medium is increased.

Amoeba↗
PubMed2004

Involvement of raft-like plasma membrane domains of Entamoeba histolytica in pinocytosis and adhesion.

Lipid rafts are highly ordered, cholesterol-rich, and detergent-resistant microdomains found in the plasma membrane of many eukaryotic cells. These domains play important roles in endocytosis, secretion, and adhesion in a variety of cell types. The parasitic protozoan Entamoeba histolytica, the causative agent of amoebic dysentery, was determined to have raft-like plasma membrane domains by use of fluorescent lipid analogs that specifically partition into raft and nonraft regions of the membrane. Disruption of raft-like membrane domains in Entamoeba with the cholesterol-binding agents filipin and methyl-beta-cyclodextrin resulted in the inhibition of several important virulence functions, fluid-phase pinocytosis, and adhesion to host cell monolayers. However, disruption of raft-like domains did not inhibit constitutive secretion of cysteine proteases, another important virulence function of Entamoeba. Flotation of the cold Triton X-100-insoluble portion of membranes on sucrose gradients revealed that the heavy, intermediate, and light subunits of the galactose-N-acetylgalactosamine-inhibitible lectin, an important cell surface adhesion molecule of Entamoeba, were enriched in cholesterol-rich (raft-like) fractions, whereas EhCP5, another cell surface molecule, was not enriched in these fractions. The subunits of the lectin were also observed in high-density, actin-rich fractions of the sucrose gradient. Together, these data suggest that pinocytosis and adhesion are raft-dependent functions in this pathogen. This is the first report describing the existence and physiological relevance of raft-like membrane domains in E. histolytica.

Animals↗
PubMed1983

Interferon treatment inhibits pinocytosis.

Treating mouse L cells with crude or purified mouse interferon inhibited fluid-phase pinocytosis. Inhibition was maximum at 24 h after treatment with 1,000 U of interferon per ml and was dose dependent and reversible with time. Pinocytosis was inhibited when human and chicken embryo cells were treated with homologous, but not heterologous, interferons.

Animals↗
PubMed1976

Contact inhibition of pleiotropin pinocytosis as a critical factor in metazoan cell ageing.

For the biosynthesis of macromolecules in amounts sufficient for indefinite growth or survival in dividing as well as in nondividing metazoan cells, a specific serum protein is required. As the addition of this factor to the medium triggers off a chain of events which leads to RNA, DNA, and protein synthesis, we have called it the pleiotropin. Pleiotropin is taken into the cells by pinocytosis. Pinocytic uptake of pleiotropin is strongly inhibited by a mutual cell-to-cell contact; diploid cells in the stage of contact inhibition of pleiotropin pinocytosis are nutritionally limited, and some changes in life processes may be expected. In continouous culture and in tissues in vivo, these changes may accumulate and ultimately result in an irreversible damage of cells so that old cells are characterized by a low pinocytic activity and by a reduced formation of macromolecules. As pleiotropin increases the saturation density and prolongs the life span of diploid cells in vitro, it is suggested that the loss of division potential in metazoan cells is not a programmed event at the cellular level and that the ageing of static cell populations may be caused by the same nutritional deficiency as that of proliferating cells.

Aging↗
PubMed1981

Adaptations of phagocytosis and pinocytosis in mouse lung macrophages after sustained in vitro hypoxia.

Alterations in lung macrophage function after chronic hypoxia have not been clearly defined. In the present studies, we examined the effect of in vitro O2 depletion (PO2 approximately 15 mmHg) for as long as 96 h on lung macrophage endocytosis. In addition, we compared the effect of an acute decrease in O2 availability on endocytic function in lung macrophages maintained for 96 h under either aerobic or hypoxic conditions. Chronic hypoxia did not result in a decreased capacity for phagocytosis. In addition, in contrast to air-maintained cells, lung macrophages exposed to low PO2 for 96 h showed no impairment in phagocytic function during acute O2 depletion. Chronic hypoxia did produce a reversible impairment in pinocytosis. However, as with phagocytosis, pinocytosis in lung macrophages pre-exposed to low PO2 for 96 h was not decreased by acute hypoxia. In these in vitro studies, chronic hypoxia appeared to produce lung macrophage adaptations that served to maintain function, despite severe O2 depletion. These adaptations may be important with respect to pinocytic and phagocytic function in clinical conditions associated wtih sustained alveolar hypoxia.

Anaerobiosis↗
PubMed1974

Pinocytosis by human alveolar macrophages. Comparison of smokers and nonsmokers.

Alveolar macrophages were obtained from human volunteers, smokers and nonsmokers, by bronchial lavage through a fiberoptic bronchoscope. Cells were incubated in a chemically defined medium containing [(14)C]sucrose (0.36 mM) and varying concentrations of rabbit serum. Pinocytosis was assessed by the cellular uptake of isotope over 30, 75, and 120-min periods. Pinocytic activity of smokers' cells was dependent on serum concentration but always less than the activity of nonsmokers' cells. The degree of pinocytosis by nonsmokers' cells was independent of serum concentration. It is concluded that the decreased level of pinocytic activity in smokers' alveolar macrophages as indicated by the uptake of sucrose in the presence of rabbit serum may represent a form of reticuloendothelial blockade.

Acid Phosphatase↗
PubMed1984

Influence of hemodynamic forces on vascular endothelial function. In vitro studies of shear stress and pinocytosis in bovine aortic cells.

The relationships between fluid shear stress, a physiologically relevant mechanical force in the circulatory system, and pinocytosis (fluid-phase endocytosis) were investigated in cultured bovine aortic endothelial cells using a specially designed apparatus. Continuous exposure to steady shear stresses (1-15 dyn/cm2) in laminar flow stimulated time- and amplitude-dependent increases in pinocytotic rate which returned to control levels after several hours. After 48 h continuous exposure to steady shear stress, removal to static conditions also resulted in a transient increase in pinocytotic rate, suggesting that temporal fluctuations in shear stress may influence endothelial cell function. Endothelial pinocytotic rates remained constant during exposure to rapidly oscillating shear stress at near physiological frequency (1 Hz) in laminar flow. In contrast, however, a sustained elevation of pinocytotic rate occurred when cells were subjected to fluctuations in shear stress amplitude (3-13 dyn/cm2) of longer cycle time (15 min), suggesting that changes in blood flow of slower periodicity may influence pinocytotic vesicle formation. As determined by [3H]thymidine autoradiography, neither steady nor oscillating shear stress stimulated the proliferation of confluent endothelial cells. These observations indicate that: (a) alterations in fluid shear stress can significantly influence the rate of formation of pinocytotic vesicles in vascular endothelial cells, (b) this process is force- and time-dependent and shows accommodation, (c) certain patterns of fluctuation in shear stress result in sustained elevation of pinocytotic rate, and (d) shear stresses can modulate endothelial pinocytosis independent of growth stimulation. These findings are relevant to (i) transendothelial transport and the metabolism of macromolecules in normal endothelium and (ii) the role of hemodynamic factors in the localization of atherosclerotic lesions in vivo.

Animals↗
PubMed1982

Redistribution of material labelled with [3H]mannose in amoebae induced to undergo pinocytosis.

The synthesis, transport, and disposition of material labelled with [3H]mannose were studied by electron microscopic radioautography in normal amoebae and in cells that had internalized cell surface as a result of being induced to undergo pinocytosis. Control amoebae were injected with the precursor and placed in normal medium. The Golgi apparatus and rough endoplasmic reticulum were heavily labelled at the earliest intervals, while radioactivity of the cell surface peaked 12 h after injection of precursor. The experimental cells were injected, placed in bovine serum albumin solution from 15 to 60 min after injection, and then removed to normal medium until fixation. Incorporation of the precursor into the rough endoplasmic reticulum was near normal, but the proportions of grains associated with the Golgi apparatus and the cell surface were greatly reduced. The percentage of grains overlying vacuoles increased 12 h after injection, notably in the case of polymorphous vacuoles and dense vacuoles, both of which were identified as lysosomes with the acid phosphatase reaction. The results suggest that addition to the surface of components labelled with [3H]mannose was diminished following induction of pinocytosis. Incorporation of the precursor appeared to be shifted from cell surface material to lysosomal contents, possibly lysosomal enzymes. It is thought that this shift occurred in response to the need for the cell to digest unusually large amounts of endocytosed protein. Recycling of cell surface under these conditions is considered possible.

Acid Phosphatase↗
PubMed1983

Coated pits with pinocytosis in Tetrahymena.

The possible role in pinocytosis of coated pits at the parasomal sacs of Tetrahymena has been studied using cationized ferritin (CF; pI = 8.5) as a marker of membrane and content. It is shown that CF binds evenly to the surface, including the coated pits, of Tetrahymena in an inorganic salt medium (to avoid formation of food vacuoles) at the normal growth temperature. Moreover, CF is internalized by coated vesicles (shown to be truly free by thin serial-section analysis) and delivered initially (1-5 min of incubation) to cisterna near the cell surface. Later (5-10 min) CF occurs also in autophagic vacuoles, formed as a result of starvation, and eventually (15-90 min) it is present in preformed (old) food vacuoles. These observations indicate that the coated pits at the parasomal sacs of Tetrahymena function in adsorptive pinocytosis in much the same manner as coated pits at the surface of mammalian cells.

Animals↗
PubMed1990

The protein kinase C inhibitor H-7 activates human neutrophils: effect on shape, actin polymerization, fluid pinocytosis and locomotion.

The present study demonstrates new properties of H-7. The protein kinase inhibitor H-7 is a potent activator of several neutrophil functions. Stimulation of initially spherical nonmotile neutrophils elicits vigorous shape changes within a few seconds, increases in cytoskeletal actin, altered F-actin distribution, increased adhesiveness and a relatively small increase in pinocytic activity. H-7 has also chemokinetic activities. Depending on the experimental condition, H-7 may elicit or inhibit neutrophil locomotion. It failed to induce chemotaxis. Thus, the response pattern elicited by H-7 is different from that of other leukocyte activators such as chemotactic peptides, PMA or diacylglycerols. The finding that H-7 can elicit shape changes, actin polymerization and pinocytosis suggests that these events can occur without activation of protein kinase C (PKC). PMA-induced shape changes and stimulation of pinocytosis were not inhibited by H-7.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗