Morphometric analysis of volumes and surface areas in membrane compartments during endocytosis in Acanthamoeba.

Bowers, Blair; Olszewski, Thomas E.; Hyde, James M.

doi:10.1083/jcb.88.3.509

Cited by 31 publications

(22 citation statements)

References 18 publications

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“…Utilizing antibodies raised against the two LPG forms, we have established the presence of LPG in internal vacuolar membranes which appear to be in equilibrium with the plasma membrane (5). We found no difference in the distribution of the two LPG forms.…”

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confidence: 74%

Localization of lipophosphonoglycan in membranes of Acanthamoeba by using specific antibodies.

Bailey¹,

Bowers²

1981

Mol. Cell. Biol.

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Antibodies against two electrophoretically distinct forms of lipophosphonoglycan (LPG) were produced in rabbits. Antibody specificity was demonstrated by the coupled antibody '25I-protein A assay (Adair et al., J. Cell Biol. 79:281-285, 1978). Indirect immunofluorescent labeling of intact Acanthamoeba showed that antibodies to both LPG components had the same uniform distribution on the cell surface. Both antibodies also bound to the cytoplasmic surface of isolated phagosomes. The location of LPG in other membranes of the amoeba was demonstrated on sections by the unlabeled antibody method. Although LPG was absent from the nuclear membrane, virtually all of the internal vacuole membranes were labeled, including the contractile vacuole. Antibodies directed against LPG were utilized to label lipophosphonoglycan in the plasma membrane of living amoebae. Labeled membrane was internalized and then localized by immunofluorescence in cytoplasmic vacuoles within 10 min of incubation. Although these results are evidence for exchange between plasma and cytoplasmic vacuolar membranes, the contractile vacuole remained unlabeled and can be considered, therefore, a separate membrane compartment. Concanavalin A also was bound and internalized by the amoeba, but electron microscopy showed that this label caused pronounced membrane perturbation, limiting its usefulness as a membrane marker in this system.

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confidence: 74%

Localization of lipophosphonoglycan in membranes of Acanthamoeba by using specific antibodies.

Bailey¹,

Bowers²

1981

Mol. Cell. Biol.

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“…14, 15 We assumed that a macrophage has an initial cell volume (V 0 ) and a total cytoplasmic membrane area (S 0 ) available for phagocytosis. For a particle with volume V p and surface area S p , the maximum number of particles that can be phagocytosed is denoted as n. After the phagocytotic process, the remaining cytoplasmic membrane area is S r and the new cell volume is V:…”

Section: Calculation Of Phagocytic Capacitymentioning

confidence: 99%

The Potential Role of Phagocytic Capacity in the Osteolytic Process Induced by Polyethylene Wear Particles

et al. 2006

J Int Med Res

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Osteolysis induced by ultra-high molecular weight polyethylene wear particles is the major cause of long-term failure of artificial joints. We examined the effects of wear particles on bioactivity by analysing the biophysical aspects of particle phagocytosis. We estimated the maximum number of internalized particles (the phagocytic capacity) for particles of various sizes and shapes. We demonstrated that elongated particles had a smaller phagocytic capacity than spherical particles of the same volume.A review of the literature showed that the ratio of particle concentration (number of particles/number of cells) to phagocytic capacity is critical for particle-induced biological responses. When this ratio was < 1, the biological response was approximately proportional to the ratio itself. When this ratio was > 1, limited changes in the biological response were observed. The saturation level of the phagocytic capacity for a particle population appears to reflect the degree of polyethylene particle-induced biological response.

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“…This compartment is maintained by the cell at virtually constant volume and surface area under standard culture conditions (3). Nevertheless, considerable membrane flow occurs through the compartment because of endocytosis (3,4). Membrane flow, as inferred from the rate of membrane ingestion by endocytosis, is rapid during pinocytosis (~9 surface equivalents/h) and close to zero in cells that have taken up particles to the limit of their capacity (that is, are saturated).…”

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Acanthamoeba discriminates internally between digestible and indigestible particles.

Bowers¹,

Olszewski²

1983

The Journal of Cell Biology

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The capacity of Acanthamoeba to distinguish nutritive yeast particles from nonnutritive plastic beads during phagocytosis was investigated. When cells were allowed to phagocytose yeast to capacity, endocytosis stopped and subsequent presentation of particles (either yeast or beads) did not result in further uptake. By contrast, when cells were allowed to phagocytose plastic beads to capacity and a second dose of particles was presented (either yeast or beads), the cells exocytosed the internal particles and took up new ones. Yeast rendered indigestible by extensive chemical cross-linking were taken up at rates similar to those of untreated yeast, but, like beads, they were exocytosed when a second dose of particles was presented. The results show that an internal distinction is made between vacuoles containing yeast and vacuoles containing plastic beads, and they are consistent with the hypothesis that the presence within the vacuoles of material capable of being digested prevents exocytosis.Phagocytic uptake of panicles depends on a specific interaction at the cell surface which results in a localized stimulus for the cell to engulf the particle. In many cells this interaction can occur with a number of indigestible substances. For example, amebas will ingest silica, carbon particles, plastic beads, and other materials of no nutritive value (1, 2). Plastic beads, in fact, are valuable tools for studying the kinetics and mechanisms of endocytosis in a wide variety of cell types because they are indiscriminately engulfed by most cells. To understand better how cells control particle turnover rates and the related membrane recycling, we have studied how Acanthamoeba handles a nutritive particle, yeast, as compared with a nonnutritive particle, plastic bead.Acanthamoeba contains a vacuolar compartment that serves as its digestive system. This compartment is maintained by the cell at virtually constant volume and surface area under standard culture conditions (3). Nevertheless, considerable membrane flow occurs through the compartment because of endocytosis (3, 4). Membrane flow, as inferred from the rate of membrane ingestion by endocytosis, is rapid during pinocytosis (~9 surface equivalents/h) and close to zero in cells that have taken up particles to the limit of their capacity (that is, are saturated). When the cell is saturated with a digestible particle such as yeast, space for particle uptake gradually becomes available as digestion progresses and undigested residues are incorporated into defecation vacuoles to be expelled from the cell. In contrast, indigestible particles like plastic beads are ingested readily by the amebas, but cannot be reduced in volume. In the experiments reported here, in addition to testing turnover rates of different types of particles, we examined whether Acanthamoeba discriminates between inert and nutritive particles and whether they do so externally or internally. MATERIALS AND METHODSCulture Methods: Acanthamoeba castellanii (Neff strain) was cultured axenically as fol...

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Morphometric analysis of volumes and surface areas in membrane compartments during endocytosis in Acanthamoeba.

Cited by 31 publications

References 18 publications

Localization of lipophosphonoglycan in membranes of Acanthamoeba by using specific antibodies.

Localization of lipophosphonoglycan in membranes of Acanthamoeba by using specific antibodies.

The Potential Role of Phagocytic Capacity in the Osteolytic Process Induced by Polyethylene Wear Particles

Acanthamoeba discriminates internally between digestible and indigestible particles.

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