Rhodamine-phalloidin staining of F-actin in pollen after dimethylsulphoxide permeabilization

Springer Science and Business Media LLC - Tập 1 - Trang 83-87 - 1988
Elisabeth S. Pierson1
1Department of Experimental Botany, Research Group Molecular Plant Physiology, University of Nijmegen, Nijmegen, The Netherlands

Tóm tắt

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.

Tài liệu tham khảo

Cresti M, Ciampolini F, Kapil RN (1984) Generative cells of some angiosperms with particular emphasis on their microtubules. J Submicrosc Cytol 16:317–326 Derksen J, Pierson ES, Traas JA (1985) Microtubules in vegetative and generative cells of pollen tubes. Eur J Cell Biol 38:142–148 Franke WW, Herth W, Van der Woude WJ, Morré DJ (1972) Tubular and filamentous structures in pollen tubes: possible involvement as guide elements in protoplasmic streaming and vectorial migration of secretory vesicles. Planta 105:317–341 Gunning BES, Hardham AR (1982) Microtubules. Annu Rev Plant Physiol 33:651–698 Heslop-Harrison J, Heslop-Harrison Y, Cresti M, Tiezzi A, Ciampolini F (1986) Actin during pollen germination. J Cell Sci 86:1–8 Heuser J, Kirschner M (1980) Filament organization revealed in platinum replicas of freeze-dried cytoskeleton. J Cell Biol 86:212–234 Lancelle SA, Cresti M, Hepler PK (1987) Ultrastructure of the cytoskeleton in freeze-substituted pollen tubes of Nicotiana alata. Protoplasma 140:141–150 Lloyd CW (1987) The plant cytoskeleton. Annu Rev Plant Physiol 38:119–139 Maupin P, Pollard T (1983) Improved preservation and staining of HeLa cell actin filaments, clathrin coated membranes and other cytoplasmic structures by tannic acid glutaralde-hyde-saponin fixation. J Cell Biol 96:51–62 Miki-Hirosige H, Nakamura S (1982) Process of metabolism during pollen tube wall formation. J Electron Microsc 31:51–62 Perdue TD, Parthasarathy MV (1985) In situ localization of F-actin in pollen tubes. Eur J Cell Biol 39:13–20 Picton JM, Steer MW (1982) A model for the mechanism of tip extension in pollen tubes. J Theor Biol 98:15–20 Pierson ES, Derksen J, Traas JA (1986) Organization of microfilaments and microtubules in pollen tubes grown in vitro or in vivo in various angiosperms. Eur J Cell Biol 41:14–18 Sanger JM, Mittal B, Pochapin M, Sanger JW (1986) Dynamics of microfilaments bundles. J Cell Sci [Suppl] 5:17–44 Tiezzi A, Cresti M, Ciampolini F (1986) Microtubules in Nicotiana pollen tubes: ultrastructural, immunofluorescence and biochemical data. In: Cresti M, Dallai R (eds) Biology of the reproduction and cell motility in plants and animals. University of Siena, Siena, pp 87–94 Tiwari SC, Wick SM, Williamson RE, Gunning BES (1984) Cytoskeleton and integration of cellular function in cells of higher plants. J Cell Biol 99:63s-69s Traas JA, Doonan JH, Rawlins D, Shaw PJ, Watts J, Lloyd CW (1987) An actin network is present in the cytoplasm throughout the division cycle of carrot cells: actin co-distributes with the four microtubule arrays. J Cell Biol 105:387–395 Wieland T (1977) Modification of actins by phallotoxins. Naturwissenschaften 64:303–309