Monitoring farnesol-induced toxicity in tobacco BY-2 cells with a fluorescent analog.
In a previous study (A. Hemmerlin, T.J. Bach, Plant Physiol. 123 (2000) 1257-1268), we have demonstrated that above a critical concentration, treatment with all-trans-farnesol induces cell-death in Nicotiana tabacum L. cv Bright Yellow-2 (TBY-2) cells. Now we used a fluorescent analog of farnesol (Fol(FLUO)), in which an isoprene unit is replaced by the fluorochrome 7-nitrobenz-2-oxa-1,3-diazol-4-yl, to visualize how cell integrity is affected. Fol(FLUO) exhibited the same toxicity as the natural compound and was shown to be readily taken up by TBY-2 cells, followed by integration into subcellular membrane structures. Although the plasma membrane seemed not to be labeled, Fol(FLUO) was associated with the tonoplast, endoplasmic reticulum, and Golgi apparatus or lipid bodies. Longer exposure times and increased Fol(FLUO) accumulation triggered the formation and proliferation of new membrane structures of as yet unknown function. Finally, at even higher and clearly cytotoxic concentrations of the analog, the cell contents became clearly disorganized, with cell swelling and ultimately plasmolysis.
Hemmerlin
Andréa
A
C.N.R.S., Institut de Biologie Moléculaire des Plantes, Département Isoprénoïdes, Université Louis Pasteur, 28 rue Goethe, F-67083 Strasbourg, France.
Reents
Reinhard
R
Mutterer
Jérôme
J
Feldtrauer
Jean-François
JF
Waldmann
Herbert
H
Bach
Thomas J
TJ
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005204:Farnesol / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D005456:Fluorescent Dyes; D008657:Metabolic Clearance Rate; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*
D005456:Fluorescent Dyes; D005204:Farnesol