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Science 299 (5604): 247-251

Copyright © 2003 by the American Association for the Advancement of Science

Regulation of Blood and Lymphatic Vascular Separation by Signaling Proteins SLP-76 and Syk

Farhad Abtahian,1* Anastasia Guerriero,1* Eric Sebzda,2 Min-Min Lu,2 Rong Zhou,3 Attila Mocsai,4 Erin E. Myers,1 Bin Huang,2 David G. Jackson,5 Victor A. Ferrari,2 Victor Tybulewicz,6 Clifford A. Lowell,4 John J. Lepore,2 Gary A. Koretzky,17dagger Mark L. Kahn2dagger

Lymphatic vessels develop from specialized endothelial cells in preexisting blood vessels, but the molecular signals that regulate this separation are unknown. Here we identify a failure to separate emerging lymphatic vessels from blood vessels in mice lacking the hematopoietic signaling protein SLP-76 or Syk. Blood-lymphatic connections lead to embryonic hemorrhage and arteriovenous shunting. Expression of slp-76 could not be detected in endothelial cells, and blood-filled lymphatics also arose in wild-type mice reconstituted with SLP-76-deficient bone marrow. These studies reveal a hematopoietic signaling pathway required for separation of the two major vascular networks in mammals.

1 Signal Transduction Program, Abramson Family Cancer Research Institute;
2 Division of Cardiology and Department of Medicine;
3 Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
4 Department of Laboratory Medicine, University of California, San Francisco, CA 94143-0134, USA.
5 MRC Human Immunology Unit, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DS, UK.
6 Division of Immune Cell Biology, National Institute for Medical Research, Mill Hill, London NW7 1AA, UK.
7 Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
*   These authors contributed equally to this work.

dagger    To whom correspondence should be addressed. E-mail: koretzky{at}mail.med.upenn.edu or markkahn{at}mail.med.upenn.edu

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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »

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