Adriana Jelínková*, Kateřina Malínská*, Sibu Simon, Juergen Kleine-Vehn, Markéta Pařezová, Přemysl Pejchar, Martin Kubeš, Jan Martinec, Jiří Friml, Eva Zažímalová and Jan Petrášek
PLANT JOURNAL
61:
883-892,
2010
Keywords:
FM 4-64, FM 1-43, plasma membrane, tobacco BY-2 cells, endocytosis
Abstract:
Remarkable progress in various techniques of in vivo fluorescence microscopy has brought an urgent need for
reliable markers for tracking cellular structures and processes. The goal of this manuscript is to describe
unexplored effects of the FM (Fei Mao) styryl dyes, which are widely used probes that label processes of
endocytosis and vesicle trafficking in eukaryotic cells. Although there are few reports on the effect of styryl
dyes on membrane fluidity and the activity of mammalian receptors, FM dyes have been considered as reliable
tools for tracking of plant endocytosis. Using plasma membrane-localized transporters for the plant hormone
auxin in tobacco BY-2 and Arabidopsis thaliana cell suspensions, we show that routinely used concentrations
of FM 4-64 and FM 5-95 trigger transient re-localization of these proteins, and FM 1-43 affects their activity. The
active process of re-localization is blocked neither by inhibitors of endocytosis nor by cytoskeletal drugs. It
does not occur in A. thaliana roots and depends on the degree of hydrophobicity (lipophilicity) of a particular
FM dye. Our results emphasize the need for circumspection during in vivo studies of membrane proteins
performed using simultaneous labelling with FM dyes.
Fulltext: contact IEB authors
IEB authors: Adriana Jelinkova,
Martin Kubeš,
Kateřina Malínská,
Jan Martinec,
Přemysl Pejchar,
Jan Petrášek,
Eva Zažímalová
Čeština
English