help button home button Biophys. J.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Biophys. J. BioFAST: First Published January 13, 2006. doi:10.1529/biophysj.105.073692
© 2006 by the Biophysical Society.

A more recent version of this article appeared on April 1, 2006.
This Article
Right arrow Full Text (Rapid PDF)
Right arrow All Versions of this Article:
biophysj.105.073692v1
90/7/2404    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Frankel, D. J.
Right arrow Articles by Burns, A. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Frankel, D. J.
Right arrow Articles by Burns, A. R.

MEMBRANES

Revealing the topography of cellular membrane domains by combined AFM/fluorescence imaging

Daniel J. Frankel 1, Janet R. Pfeiffer 2, Zurab Surviladze 2, A. E. Johnson 3, Janet M Oliver 2, Bridget S. Wilson 2 and Alan R. Burns 1*

1 Sandia National Laboratories
2 Univ. of New Mexico
3 Texas A&M Univ.

* To whom correspondence should be addressed. E-mail: aburns{at}sandia.gov.

Submitted on August 31, 2005
Revised on October 3, 2005
Accepted on 14 December 2005


  Abstract
Simultaneous atomic force microscopy (AFM) and confocal fluorescence imaging was used to observe in aqueous buffer the 3-dimensional landscape of the inner surface of membrane sheets stripped from fixed tumor mast cells. The AFM images reveal prominent, irregularly shaped raised domains that label with fluorescent markers for both resting and activated IgE receptors (Fc{epsilon}RI), as well as with cholera toxin-aggregated GM1 and clathrin. The latter suggests that coated pits bud from these regions. These features are interspersed with flatter regions of membrane and are frequently surrounded and interconnected by cytoskeletal assemblies. The raised domains shrink in height by roughly 50% when cholesterol is extracted with methyl-{beta}-cyclodextrin. Based on composition, the raised domains seen by AFM correspond to the cholesterol-enriched dark patches observed in transmission electron microscopy (TEM). These patches were previously identified as sites of signaling and endocytosis based on their localization of activated Fc{epsilon}RI, at least 10 associated signaling molecules, and the presence of clathrin-coated pits (1). Overall the data suggest that signaling and endocytosis occur in mast cells from raised membrane regions that depend on cholesterol for their integrity and may be organized in specific relationship with the cortical cytoskeleton.

Key Words: AFM, IgE receptor, TEM, domains, membranes




This article has been cited by other articles:


Home page
 Antimicrob. Agents Chemother.Home page
J. C. Fernandes Rodrigues, J. L. Concepcion, C. Rodrigues, A. Caldera, J. A. Urbina, and W. de Souza
In Vitro Activities of ER-119884 and E5700, Two Potent Squalene Synthase Inhibitors, against Leishmania amazonensis: Antiproliferative, Biochemical, and Ultrastructural Effects
Antimicrob. Agents Chemother., November 1, 2008; 52(11): 4098 - 4114.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
A. Yersin, H. Hirling, S. Kasas, C. Roduit, K. Kulangara, G. Dietler, F. Lafont, S. Catsicas, and P. Steiner
Elastic Properties of the Cell Surface and Trafficking of Single AMPA Receptors in Living Hippocampal Neurons
Biophys. J., June 15, 2007; 92(12): 4482 - 4489.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Copyright © 2006 by the Biophysical Society.