Skip Navigation


Plant and Cell Physiology Advance Access originally published online on August 29, 2008
Plant and Cell Physiology 2008 49(10):1508-1521; doi:10.1093/pcp/pcn122
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplementary Data
Right arrow All Versions of this Article:
49/10/1508    most recent
pcn122v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Klima, A.
Right arrow Articles by Foissner, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Klima, A.
Right arrow Articles by Foissner, I.
Agricola
Right arrow Articles by Klima, A.
Right arrow Articles by Foissner, I.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

FM Dyes Label Sterol-Rich Plasma Membrane Domains and are Internalized Independently of the Cytoskeleton in Characean Internodal Cells

Andreas Klima and Ilse Foissner*

Department of Cell Biology, Division of Plant Physiology, University of Salzburg, Hellbrunnerstrasse 34, A-5020 Salzburg, Austria

*Corresponding author: E-mail, ilse.foissner{at}sbg.ac.at; Fax, +43-662-8044-619.


  Abstract

We applied the endocytic markers FM1-43, FM4-64 and filipin to internodal cells of the green alga Chara corallina. Both FM dyes stained stable, long-living plasma membrane patches with a diameter of up to 1 µm. After 5 min, FM dyes labeled cortical, trembling structures up to 500 nm in size. After 15 min, FM dyes localized to endoplasmic organelles up to 1 µm in diameter, which migrated actively along actin bundles or participated in cytoplasmic mass streaming. After 30–60 min, FM fluorescence appeared in the membrane of small, endoplasmic vacuoles but not in that of the central vacuole. Some of the FM-labeled organelles were also stained by neutral red and lysotracker yellow, indicative of acidic compartments. Filipin, a sterol-specific marker, likewise labeled plasma membrane domains which co-localized with the FM patches. However, internalization of filipin could not be observed. KCN, cytochalasin D, latrunculin B and oryzalin had no effect on size, shape and distribution of FM- and filipin-labeled plasma membrane domains. Internalization of FM dyes was inhibited by KCN but not by drugs which interfere with the actin or microtubule cytoskeleton. Our data indicate that the plasma membrane of characean internodal cells contains discrete domains which are enriched in sterols and probably correspond to clusters of lipid rafts. The inhibitor experiments suggest that FM uptake is active but independent of actin filaments, actin polymerization and microtubules. The possible function of the sterol-rich, FM labeled plasma membrane areas and the significance of actin-independent FM internalization (via endocytosis or energy-dependent flippases) are discussed.

Keywords: Chara - Cytoskeleton - Endocytosis - Lipid raft - Plasma membrane domain - Sterol

Abbreviations: AFW, artificial fresh water; BDM, 2,3-butanedione monoxime; CD, cytochalasin D; CLSM, confocal laser scanning microscope/microscopy; DIC, differential interference contrast; DiOC6(3), 3,3', dihexyloxacarbocyanine iodide; DMSO, dimethylsulfoxide; FRAP, fluorescence recovery after photobleaching; LatB, latrunculin B; PBS, phosphate-buffered saline; PIPES, piperazine-N,N'-bis (2-ethanesulfonic acid).

(Received July 21, 2008; Accepted August 14, 2008)
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.