Myosin reorganization in activated RBL cells correlates temporally with stimulated secretion.
Rat basophilic leukemia cells secrete histamine and serotonin in response to cross-linking of the IgE receptor by multivalent antigen [Metzger et al., 1986: Ann. Rev. Immunol. 4:419-470]. Receptor crosslinking also induces phosphorylation of the light and heavy chains of myosin II with kinetics similar to that of secretion [Ludowyke et al., 1989: J. Biol. Chem. 264:12492-12501]. Here we show that myosin II localization changes after activation with similar kinetics. Furthermore, these changes are coincident with changes in cell shape and increase in motile activity induced by activation. Within 2 min, activated cells begin to flatten, spread on their substratum, and extend lamellipodia which show active ruffling. Quantitation of the extent of cell spreading from video micrographs shows that 48% of the cells increase significantly in surface area by 5 min and 71% by 15 min. Myosin II is uniformly distributed in unactivated cells but is deficient in newly formed lamellipodia that start to appear at 2 min after activation. In contrast these lamellipodia show strong staining for actin. Further changes in myosin organization are detected by 15 min after activation when myosin reappears in the cell periphery, is concentrated in the perinuclear area, and is also organized in punctate linear arrays that extend from the nucleus to the cell periphery. The kinetics of the early cell shape changes and formation of the myosin-deficient lamellipodia correlate well with, and may relate to, the increase in the level of myosin II phosphorylation reported by Ludowyke et al. [1989: J. Biol. Chem. 264:12492-12501]. Changes in the distribution of cell surface-bound IgE also occur upon antigen activation, and they correlate with the myosin distribution in a manner that suggests that they may be driven by myosin II.
Spudich
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Department of Molecular and Cellular Physiology, Stanford University School of Medicine, California 94305.
D000199:Actins / Q000032:analysis / Q000648:ultrastructure; D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure; D002465:Cell Movement; D048429:Cell Size; D007073:Immunoglobulin E / Q000378:metabolism / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute; D008407:Mast Cells / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009218:Myosins / Q000032:analysis* / Q000648:ultrastructure; D010590:Phalloidine / Q000494:pharmacology; D051381:Rats; D017455:Receptors, IgE / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000199:Actins; D017455:Receptors, IgE; D010590:Phalloidine; D007073:Immunoglobulin E; D009218:Myosins