| Investigating cellular signaling reactions in single attoliter vesicles. | |
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MedLine Citation:
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PMID: 15740126 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Horst Pick; Evelyne L Schmid; Ana-Paula Tairi; Erwin Ilegems; Ruud Hovius; Horst Vogel |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of the American Chemical Society Volume: 127 ISSN: 0002-7863 ISO Abbreviation: J. Am. Chem. Soc. Publication Date: 2005 Mar |
Date Detail:
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Created Date: 2005-03-02 Completed Date: 2005-04-20 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 7503056 Medline TA: J Am Chem Soc Country: United States |
Other Details:
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Languages: eng Pagination: 2908-12 Citation Subset: IM |
Affiliation:
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Laboratory of Physical Chemistry of Polymers and Membranes, Swiss Federal Institute of Technology in Lausanne (EPFL), 1015 Ecublens, Switzerland. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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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:
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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 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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