Proceedings of the National Academy of Sciences · 1980 · 443 citations · 17 references
Life SciencesBiologyCell BiologyCytoskeletonMolecular Cell BiologyPhycologyActin FilamentsAlgal Cell BiologyCytoplasmic StreamingCytoskeletal Associated ProteinsCytoskeletal DynamicsCytoskeleton DynamicsActin Cytoskeleton DynamicsActin DynamicsActin-binding ProteinsAlgal Cytoplasmic StreamingCompetitive Binding AssaysCompetitive Binding StudiesFluorescent LabelingStress Fibers
An active fluorescent derivative of the actin-binding mushroom toxin phallacidin has been synthesized. Convenient methods were developed to stain actin cytoskeletal structures in living and fixed cultured animal cells and actively streaming algal cells. Actin binding specificity was demonstrated by competitive binding experiments and comparative staining of well-known structures. Large populations of living animal cells in culture were readily stained by using a relatively mild lysolecithin permeabilization procedure facilitated by the small molecular size of the label. Actin in animal cells was stained stress fibers, ruffles, the cellular geodome, and in diffuse appearing distributions apparently associated with the plasma membrane. Staining of actin cables in algae with nitrobenzoxadiazole (NBD)-phallacidin did not inhibit cytoplasmic streaming. NBD-phallacidin provides a convenient actin-specific fluorescent label for cellular cytoskeletal structures with promise for use in studies of actin dynamics in living systems.
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