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Chemoattractant-elicited translocation of myosin in motileDictyostelium by Vivianne T. Nachmias; Yoshio Fukui; James A. Spudich is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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The distribution of myosin was studied in amebae of the Ax-3 and NC-4 strains of Dictyosteliurn migrating at room temperature, using indirect immunofluorescence of aggregation-competent amebae and the agar-overlay technique. Amebae were fixed in methanol-formaldehyde or absolute acetone at -15°C before or after stimulation with micromolar cyclic AMP at room temperature (20-25°C). Myosin was detected by monoclonal antibodies to Dictyosteliurn myosin heavy chain followed by a fluorescent secondary antibody that had been preabsorbed to remove nonspecific staining. In both strains there was a striking increase in intensity of anti-myosin immunofluorescence in the cortex where it appeared as a continuous ring 30 seconds after addition of cyclic AMP. This correlated with a rounding up of the cell body. Sixty seconds after stimulation there was a clear reduction of cytoplasmic myosin rods in conjunction with the increased cortical localization. At this time extensions of largely hyaline cytoplasm were observed that extended beyond the cortical shell of myosin. Two minutes after the stimulus the immunofluorescence remained as a distinct line at the cortex, but the cells began to resume in

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Author

Vivianne T. Nachmias; Yoshio Fukui; James A. Spudich

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 0886-1544)

Published

1988

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)