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Title: Investigating cellular signaling reactions in single attoliter vesicles
Authors: Pick, H.
Schmidt, E.L. 
Tairi, A.-P.
Ilegems, E.
Hovius, R.
Vogel, H.
Issue Date: 9-Mar-2005
Citation: Pick, H., Schmidt, E.L., Tairi, A.-P., Ilegems, E., Hovius, R., Vogel, H. (2005-03-09). Investigating cellular signaling reactions in single attoliter vesicles. Journal of the American Chemical Society 127 (9) : 2908-2912. ScholarBank@NUS Repository.
Abstract: 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-HT3 receptor and subsequent transmembrane Ca2+ 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. © 2005 American Chemical Society.
Source Title: Journal of the American Chemical Society
ISSN: 00027863
DOI: 10.1021/ja044605x
Appears in Collections:Staff Publications

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