Rhodamine-phalloidin staining of F-actin in pollen after dimethylsulphoxide permeabilization |
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Authors: | Elisabeth S Pierson |
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Institution: | (1) Department of Experimental Botany, Research Group Molecular Plant Physiology, University of Nijmegen, Toernooiveld, NL-6525 ED Nijmegen, The Netherlands |
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Abstract: | Summary Bundles of actin filaments were observed in in vitro cultured pollen of Lilium longiflorum and Nicotiana tabacum which had been permeabilized in a buffered medium containing 5% dimethylsulphoxide (DMSO), EGTA, rhodaminephalloidin for F-actin staining, and sucrose as an osmoticum. In imbibed pollen grains, especially those of lily, numerous short bundles and small foci of F-actin were clearly visible. In germinated pollen grains, fine bundles of F-actin could be seen to converge at the aperture of the pollen grain. Along the entire length of the pollen tube, including the extreme tip, a dense three-dimensional netaxial distribution of actin filaments was observed. The F-actin patterns visualized after permeabilization with DMSO are much finer and more detailed than those observed after conventional fixation with formaldehyde. |
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Keywords: | Cytoskeleton Dimethylsulphoxide F-actin Pollen Rhodamine-phalloidin |
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