Document Detail


Investigating cellular signaling reactions in single attoliter vesicles.
MedLine Citation:
PMID:  15740126     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Understanding cellular signaling mediated by cell surface receptors is key to modern biomedical research and drug development. The discovery of a growing number of potential molecular targets and therapeutic compounds requires downscaling and accelerated functional screening. Receptor-mediated cellular responses are typically investigated on single cells or cell populations. Here, we show how to monitor cellular signaling reactions at a yet unreached miniaturization level. On the basis of our observations, cytochalasin induces mammalian cells to extrude from their plasma membrane submicrometer-sized native vesicles. They comprise functional cell surface receptors correctly exposing their extracellular ligand binding sites on the outer vesicle surface and retaining cytosolic proteins in the vesicle interior. As a prototypical example, ligand binding to the ionotropic 5-HT(3) receptor and subsequent transmembrane Ca(2+) signaling were monitored in single attoliter vesicles. Thus, native vesicles are the smallest autonomous containers capable of performing cellular signaling reactions under physiological conditions. Because a single cell delivers about 50 native vesicles, which can be isolated and addressed as individuals, our concept allows multiple functional analyses of individual cells having a limited availability and opens new vistas for miniaturized bioanalytics.
Authors:
Horst Pick; Evelyne L Schmid; Ana-Paula Tairi; Erwin Ilegems; Ruud Hovius; Horst Vogel
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  127     ISSN:  0002-7863     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2005 Mar 
Date Detail:
Created Date:  2005-03-02     Completed Date:  2005-04-20     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2908-12     Citation Subset:  IM    
Affiliation:
Laboratory of Physical Chemistry of Polymers and Membranes, Swiss Federal Institute of Technology in Lausanne (EPFL), 1015 Ecublens, Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Calcium Signaling / physiology*
Cell Line
Cell Membrane / chemistry,  drug effects,  metabolism
Cytochalasin B / pharmacology
Cytosol / chemistry,  metabolism
Humans
Kidney / cytology
Ligands
Mice
Microscopy, Confocal
Microscopy, Fluorescence
Receptor, Epidermal Growth Factor / chemistry,  metabolism
Receptors, Serotonin, 5-HT3 / chemistry,  metabolism,  physiology*
Recombinant Fusion Proteins / chemistry,  metabolism,  physiology
Chemical
Reg. No./Substance:
0/Ligands; 0/Receptors, Serotonin, 5-HT3; 0/Recombinant Fusion Proteins; 14930-96-2/Cytochalasin B; EC 2.7.10.1/Receptor, Epidermal Growth Factor

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