Intracellular delivery of carbohydrates into mammalian cells through swelling-activated pathways

被引:49
作者
Reuss, R
Ludwig, J
Shirakashi, R
Ehrhart, F
Zimmermann, H
Schneider, S
Weber, MM
Zimmermann, U
Schneider, H
Sukhorukov, VL
机构
[1] Univ Wurzburg, Lehrstuhl Biotechnol, Biozentrum, D-97074 Wurzburg, Germany
[2] Univ Tokyo, Inst Ind Sci, Meguro Ku, Tokyo 1538505, Japan
[3] Fraunhofer Inst Biomed Tech, Abt Kryobiophys & Kryotechnol, D-66386 St Ingbert, Germany
[4] Univ Klin Bochum, Allgemeine Innere Med Endokrinol & Diabetol, D-44789 Bochum, Germany
[5] Univ Klin Mainz, Med Klin & Poliklin 1, Schwepunkt Endokrinol & Stoffwechselerkrankungen, D-55131 Mainz, Germany
关键词
cell volumetry; regulatory volume decrease; osmotic stress; trehalose; inositol; sorbitol; Jurkat cells; T-lymphocytes; volume-sensitive channels;
D O I
10.1007/s00232-004-0694-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Volume changes of human T-lymphocytes (Jurkat line) exposed to hypotonic carbohydrate-substituted solutions of different composition and osmolality were studied by videomicroscopy. In 200 mOsm media the cells first swelled within 1-2 min and then underwent regulatory volume decrease (RVD) to their original isotonic volume within 10-15 min. RVD also occurred in strongly hypotonic 100 mOsni solutions of di- and trisaccharides (trehalose, sucrose, raffinose). In contrast to oligosaccharide media, 100 mOsm solutions of monomeric carbohydrates (glucose. galactose, mositol and sorbitol) inhibited RVD. The complex volumetric data were analyzed with a membrane transport model that allowed the estimation of the hydraulic conductivity volume-dependent Solute permeabilities and ties. We found that under slightly hypotonic stress (200 mOsm) the cell membrane was impermeable to all carbohydrates studied here. Upon osmolality decrease to 100 mOsm, the membrane permeability to monomeric carbohydrates increased dramatically (apparently due to channel activation caused by extensive cell swelling), whereas oligosaccharide permeability remained very poor. The size-selectivity of the swelling-activated sugar permeation was confirmed by direct chromatographic measurements of intracellular sugars. The results of this study are of interest for biotechnology, where sugars and related compounds are increasingly being used as potential cryo- and lyoprotective agents for preservation of rare and valuable mammalian cells and tissues.
引用
收藏
页码:67 / 81
页数:15
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