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Plant Cell. 2002 Jan;14(1):71-86.

Uptake of a fluorescent marker in plant cells is sensitive to brefeldin A and wortmannin.

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  • 1Institute for Molecular Biotechnology, Biology VII, Rheinisch-Westfälische Technische Hochschule Aachen, Worringerweg 1, D-52074 Aachen, Germany. emans@molbiotech.rwth-aachen.de

Abstract

We assessed FM1-43 [N-(3-triethylammoniumpropyl)-4-(4-[dibutylamino]styryl)pyridinium dibromide] as a fluorescent endocytosis marker in intact, walled plant cells. At 4 degrees C, FM1-43 stained the plasma membrane, and after 30 to 120 min of incubation at 26 degrees C, FM1-43 labeled cytoplasmic vesicles and then the vacuole. Fluorimetric quantitation demonstrated dye uptake temperature sensitivity (approximately 65% reduction at 16 degrees C, >90% at 4 degrees C). FM1-43 uptake in suspension cells was stimulated more than twofold by brefeldin A and inhibited approximately 0.4-fold by wortmannin. FM1-43 delivery to the vacuole was largely inhibited by brefeldin A, although overall uptake was stimulated, and brefeldin A treatment caused the accumulation of large prevacuolar endosomal vesicles heavily labeled with FM1-43. Three-dimensional time lapse imaging revealed that FM1-43-labeled vacuoles and vesicles are highly dynamic. Thus, FM1-43 serves as a fluorescent marker for imaging and quantifying membrane endocytosis in intact plant cells.

PMID:
11826300
PMCID:
PMC150552
[PubMed - indexed for MEDLINE]
Free PMC Article
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