Abstract
The phagocytic ability of amoebae of the cellular slime mold Polysphondylium pallidum, grown in shaken suspension, was examined. An established quantitative assay of the uptake of polystyrene (PS) beads was shown to be valid for this organism. The kinetics of phagocytosis were determined, and estimates of the concentration of PS beads necessary to achieve half-maximal phagocytic velocity (Kp), as well as the maximal velocity itself (Vp max), were made. Comparison with previously published data on Acanthamoeba and guinea pig leukocytes suggested that the P. pallidum amoebae had the lowest Kp, while the leukocytes had the highest Vp max. Beads approximately 1 µm in diameter appeared to be the optimal size for ingestion. Simultaneously with phagocytosis, comparable numbers of beads accumulated at the cell surface; this accumulation did not occur when phagocytosis was inhibited. Phagocytosis was depressed by protein in the medium, by increased osmolarity, and by inhibitors of aerobic metabolism. Starvation-initiated development, leading to encystment, was shown to affect the capacity of the cells to phagocytize, mainly by progressively decreasing the time span over which the cells ingested particles at a constant initial rate.
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- Allison A. C., Davies P., De Petris S. Role of contractile microfilaments in macrophage movement and endocytosis. Nat New Biol. 1971 Aug 4;232(31):153–155. doi: 10.1038/newbio232153a0. [DOI] [PubMed] [Google Scholar]
- Baehner R. L., Nathan D. G., Karnovsky M. L. Correction of metabolic deficiencies in the leukocytes of patients with chronic granulomatous disease. J Clin Invest. 1970 May;49(5):865–870. doi: 10.1172/JCI106305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowers B., Korn E. D. The fine structure of Acanthamoeba castellanii (Neff strain). II. Encystment. J Cell Biol. 1969 Jun;41(3):786–805. doi: 10.1083/jcb.41.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowers B., Olszewski T. E. Pinocytosis in Acanthamoeba castellanii. Kinetics and morphology. J Cell Biol. 1972 Jun;53(3):681–694. doi: 10.1083/jcb.53.3.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CHAPMAN-ANDRESEN C., HOLTER H. Studies on the ingestion of 14C glucose by pinocytosis in the amoeba Chaos chaos. Exp Cell Res. 1955;(Suppl 3):52–63. [PubMed] [Google Scholar]
- Davis A. T., Estensen R., Quie P. G. Cytochalasin B. 3. Inhibition of human polymorphonuclear leukocyte phagocytosis. Proc Soc Exp Biol Med. 1971 May;137(1):161–164. doi: 10.3181/00379727-137-35535. [DOI] [PubMed] [Google Scholar]
- Gerisch G. Cell aggregation and differentiation in Dictyostelium. Curr Top Dev Biol. 1968;3:157–197. doi: 10.1016/s0070-2153(08)60354-3. [DOI] [PubMed] [Google Scholar]
- Githens S., 3rd, Karnovsky M. L. Biochemical changes during growth and encystment of the cellular slime mold Polysphondylium pallidum. J Cell Biol. 1973 Sep;58(3):522–535. doi: 10.1083/jcb.58.3.522. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HOFSTEE B. H. J. On the evaluation of the constants Vm and KM in enzyme reactions. Science. 1952 Sep 26;116(3013):329–331. doi: 10.1126/science.116.3013.329. [DOI] [PubMed] [Google Scholar]
- Korn E. D., Weisman R. A. Phagocytosis of latex beads by Acanthamoeba. II. Electron microscopic study of the initial events. J Cell Biol. 1967 Jul;34(1):219–227. doi: 10.1083/jcb.34.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Michell R. H., Pancake S. J., Noseworthy J., Karnovsky M. L. Measurement of rates of phagocytosis: the use of cellular monolayers. J Cell Biol. 1969 Jan;40(1):216–224. doi: 10.1083/jcb.40.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pollard T. D., Shelton E., Weihing R. R., Korn E. D. Ultrastructural characterization of F-actin isolated from Acanthamoeba castellanii and identification of cytoplasmic filaments as F-actin by reaction with rabbit heavy meromyosin. J Mol Biol. 1970 May 28;50(1):91–97. doi: 10.1016/0022-2836(70)90106-3. [DOI] [PubMed] [Google Scholar]
- ROBERTS J., QUASTEL J. H. PARTICLE UPTAKE BY POLYMORPHONUCLEAR LEUCOCYTES AND EHRLICH ASCITES-CARCINOMA CELLS. Biochem J. 1963 Oct;89:150–156. doi: 10.1042/bj0890150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SBARRA A. J., KARNOVSKY M. L. The biochemical basis of phagocytosis. 2. Incorporation of C14-labeled building blocks into lipid, protein, and glycogen of leukocytes during phagocytosis. J Biol Chem. 1960 Aug;235:2224–2229. [PubMed] [Google Scholar]
- SBARRA A. J., KARNOVSKY M. L. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J Biol Chem. 1959 Jun;234(6):1355–1362. [PubMed] [Google Scholar]
- SONNEBORN D. R., SUSSMAN M., LEVINE L. SEROLOGICAL ANALYSES OF CELLULAR SLIME-MOLD DEVELOPMENT. I. CHANGES IN ANTIGENIC ACTIVITY DURING CELL AGGREATION. J Bacteriol. 1964 Jun;87:1321–1329. doi: 10.1128/jb.87.6.1321-1329.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ulsamer A. G., Smith F. R., Korn E. D. Lipids of Acanthamoeba castellanii. Composition and effects of phagocytosis on incorporation of radioactive precursors. J Cell Biol. 1969 Oct;43(1):105–114. doi: 10.1083/jcb.43.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WRIGHT B. E., BLOOM B. In vivo evidence for metabolic shifts in the differentiating slime mold. Biochim Biophys Acta. 1961 Apr 1;48:342–346. doi: 10.1016/0006-3002(61)90484-x. [DOI] [PubMed] [Google Scholar]
- Weihing R. R., Korn E. D. Acanthamoeba actin. Isolation and properties. Biochemistry. 1971 Feb 16;10(4):590–600. doi: 10.1021/bi00780a008. [DOI] [PubMed] [Google Scholar]
- Weisman R. A., Korn E. D. Phagocytosis of latex beads by Acanthamoeba. I. Biochemical properties. Biochemistry. 1967 Feb;6(2):485–497. doi: 10.1021/bi00854a017. [DOI] [PubMed] [Google Scholar]
- Weisman R. A., Moore M. O. Bead uptake as a tool for studying differentiation in Acanthamoeba. Exp Cell Res. 1969 Jan;54(1):17–22. doi: 10.1016/0014-4827(69)90286-9. [DOI] [PubMed] [Google Scholar]
- Wessells N. K., Spooner B. S., Ash J. F., Bradley M. O., Luduena M. A., Taylor E. L., Wrenn J. T., Yamada K. Microfilaments in cellular and developmental processes. Science. 1971 Jan 15;171(3967):135–143. doi: 10.1126/science.171.3967.135. [DOI] [PubMed] [Google Scholar]
- Woolley D. E. Extraction of an actomyosin-like pootein from amoebae of Dictyostelium discoideum. J Cell Physiol. 1970 Oct;76(2):185–190. doi: 10.1002/jcp.1040760208. [DOI] [PubMed] [Google Scholar]
- ZUCKER-FRANKLIN D., HIRSCH J. G. ELECTRON MICROSCOPE STUDIES ON THE DEGRANULATION OF RABBIT PERITONEAL LEUKOCYTES DURING PHAGOCYTOSIS. J Exp Med. 1964 Oct 1;120:569–576. doi: 10.1084/jem.120.4.569. [DOI] [PMC free article] [PubMed] [Google Scholar]