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First published online 27 January 2009
doi: 10.1242/jcs.034207

Journal of Cell Science 122, 505-512 (2009)
Published by The Company of Biologists 2009
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Research Article

Sphingosine-1-phosphate receptors stimulate macrophage plasma-membrane actin assembly via ADP release, ATP synthesis and P2X7R activation

Mark P. Kuehnel1, Miriam Reiss1, Paras K. Anand1, Irina Treede1, Daniela Holzer1, Eik Hoffmann1,2, Manuela Klapperstueck3, Thomas H. Steinberg4, Fritz Markwardt3 and Gareth Griffiths1,*

1 EMBL, Meyerhofstr. 1, 69117 Heidelberg, Germany
2 Institute for Biological Sciences, University of Rostock, Albert-Einstein-Str. 3, 18051 Rostock, Germany
3 Julius-Bernstein-Institute for Physiology, Martin-Luther-University Magdeburger Str. 6, 06097 Halle, Germany
4 Department of Internal Medicine, Washington University School of Medicine, St Louis, MO 63110, USA

* Author for correspondence (e-mail: griffiths{at}embl.de)

Accepted 23 October 2008

Eukaryotic plasma membranes assemble actin filaments within seconds of activation of many receptors, especially during chemotaxis. Here, serum or sphingosine-1-phosphate stimulation of J774 and RAW macrophages released ADP within seconds into the extracellular medium, along with an adenylate kinase activity that converted ADP to ATP. ATP then activated the P2X7 receptor (P2X7R) that was necessary for a peak of plasma-membrane actin assembly within 5 to 10 seconds in P2X7R-expressing J774, RAW and primary macrophages. Neither actin assembly nor characteristic P2X7R channel activity was seen in response to ATP in P2X7R-knockout macrophages, as detected by patch-clamp analysis. Since P2X7R has been shown previously to form a macromolecular complex with actin we propose that it is involved in the membrane assembly of actin. Our data reveal a surprisingly rapid and complex relay of signaling and externalization events that precede and control actin assembly induced by sphingosine-1-phosphate. The overall model we present is strongly supported by the data presented in the accompanying paper that focuses on latex bead phagosomes.

Key words: ATP, P2X7, Sphingosine-1-P, Actin, Macrophages, Phagosomes


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Mark. P. Kuehnel, V. Rybin, P. K. Anand, E. Anes, and G. Griffiths
Lipids regulate P2X7-receptor-dependent actin assembly by phagosomes via ADP translocation and ATP synthesis in the phagosome lumen
J. Cell Sci., February 15, 2009; 122(4): 499 - 504.
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