2Komarov Botanical Institute, Russian Academy of Sciences, St. Petersburg, Russia; E-mail: email@example.com
3Institute of Analytical Instrument Engineering, Russian Academy of Sciences, St. Petersburg, Russia; E-mail: firstname.lastname@example.org
4Uppsala University, P. O. Box 256, 75105 Uppsala, Sweden; E-mail: Roman.Zubarev@icm.uu.se
* To whom correspondence should be addressed.
Received October 2, 2009; Revision received November 25, 2009
The present work was devoted to the exploration of the role of sterols in the functioning of membranes in root cells. Membrane characteristics and composition of the membrane lipids in the roots of wheat (Triticum aestivum L.) seedlings treated with exogenous cholesterol and antibiotic nystatin, which specifically binds with endogenous sterols, were analyzed. Cholesterol caused a fall of membrane potential, acidification of the incubation medium, decrease in potassium leakage of roots, and increase in the level of exogenous superoxide radical. Similarly to cholesterol, the application of nystatin also induced the depolarization of the plasma membrane, but in contrast with cholesterol it was accompanied by alkalinization of the incubation medium and decrease in the level of exogenous superoxide radical. Analysis of membrane lipids showed that following nystatin treatment the total sterol content in roots did not change, while the level of complex sphingolipids represented mainly by glycoceramides became higher. Using mass spectrometry with electrospray ionization (+ESI-MS) for the analysis of the glycoceramide composition, we showed that nystatin induced changes in the ratios of molecular species of glycoceramides. It was suggested that the modification of the sterol component of plasma membrane could influence membrane functioning by changing the sphingolipid composition of lipid rafts.
KEY WORDS: wheat roots, nystatin, sterols, glycoceramides, reactive oxygen species, mass spectrometry with electrospray ionization