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MBC in Press, published online ahead of print January 12, 2004
Mol. Biol. Cell 10.1091/mbc.E03-06-0391

A more recent version of this article appeared on March 1, 2004
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Submitted on June 12, 2003
Revised on November 9, 2003
Accepted on November 21, 2003

Cardiac steroids induce changes in recycling of the plasma membrane in human NT2 cells

Haim Rosen1*, Vladimir Glukmann2, Tomer Feldmann2, Eleonora Fridman2, and David Lichtstein3*

1 The Kuvin Center for the Study of Infectious and Tropical Diseases, Institute of Microbiology, Department of Physiology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel
2 The Kuvin Center for the Study of Infectious and Tropical Diseases, Institute of Microbiology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel
3 Department of Physiology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel

* Corresponding author. E-mail address: david{at}md2.huji.ac.il.

* Corresponding author. E-mail address: david{at}md2.huji.ac.il.

Cardiac steroids (CS) are specific inhibitors of Na+, K+-ATPase activity. Although the presence of CS-like compounds in animal tissues has been established, their physiological role is not evident. In the present study, treatment of human NT2 cells with physiological concentrations of CS (nM) caused the accumulation of large vesicles adjacent to the nucleus. Experiments using FM1-43, transferrin, LDL and selected antitransferrin-receptor and Rab protein antibodies revealed that CS induced changes in endocytosis-dependent membrane traffic. Our data indicate that the CS-induced accumulation of cytoplasmic membrane components is a result of inhibited recycling within the late endocytic pathway. Furthermore, our results support the notion that the CS-induced changes in membrane traffic is mediated by the Na+, K+-ATPase. These phenomena were apparent in NT2 cells at nM concentrations of CS and were observed also in other human cell lines, pointing to the generality of this phenomenon. Based on these observations, we propose that the endogenous CS-like compounds are physiological regulators of recycling of endocytosed membrane proteins and cargo.




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