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ABSTRACT Human immune cells have to penetrate an endothelial barrier during their beneficial pursuit of infection and their destructive infiltration of tissues in autoimmune diseases. This

transmigration requires Rap1 GTPase to activate integrin affinity1. We define a new model system for this process by demonstrating, with live imaging and genetics, that during embryonic

development _Drosophila melanogaster_ immune cells penetrate an epithelial, _Drosophila_ E-cadherin (DE-cadherin)-based tissue barrier. A mutant in RhoL, a GTPase homologue that is

specifically expressed in haemocytes, blocks this invasive step but not other aspects of guided migration. RhoL mediates integrin adhesion caused by _Drosophila_ Rap1 overexpression and

moves Rap1 away from a concentration in the cytoplasm to the leading edge during invasive migration. These findings indicate that a programmed migratory step during _Drosophila_ development

bears striking molecular similarities to vertebrate immune cell transmigration during inflammation, and identify RhoL as a new regulator of invasion, adhesion and Rap1 localization. Our work

establishes the utility of _Drosophila_ for identifying novel components of immune cell transmigration and for understanding the _in vivo_ interplay of immune cells with the barriers they

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Download references ACKNOWLEDGEMENTS We thank D. Jukam, P. Kunwar, A-M. Sudarov, and S. Wang for help during the GAL4 enhancer screen, and D. Demy for input. We thank all the people who

shared their reagents: U. Heberlein, K. Brueckner, N. Perrimon, D. Montell, R. Reuter and A. Page-McCaw for stocks and T. Uemura, D. Branton, R, Dubreuil, H. Bellen, N. Lowe, D. Ryoo and J.

Treisman for antibodies. We thank the Developmental Studies Hybridoma Bank developed with NICHD support and maintained by the University of Iowa for antibodies and the Bloomington Stock

Center for flies. We are grateful for conversations with P. Rangan, P. Kunwar and many other members of the laboratory. We thank M. Dustin, S. Schwab and laboratory members for comments on

the manuscript. We thank NICHD for supporting part of this work. R.L. is an investigator of the HHMI. AUTHOR INFORMATION Author notes * Martin Haesemeyer Present address: Current address:

Research Institute of Molecular Pathology (IMP), Dr. Bohrgasse 7, A-1030 Vienna, Austria., AUTHORS AND AFFILIATIONS * Department of Cell Biology, HHMI and Kimmel Center for Biology and

Medicine of the Skirball Institute, New York University School of Medicine, New York, 10016-6481, New York Daria Siekhaus, Martin Haesemeyer, Olivia Moffitt & Ruth Lehmann Authors *

Daria Siekhaus View author publications You can also search for this author inPubMed Google Scholar * Martin Haesemeyer View author publications You can also search for this author inPubMed 

Google Scholar * Olivia Moffitt View author publications You can also search for this author inPubMed Google Scholar * Ruth Lehmann View author publications You can also search for this

author inPubMed Google Scholar CONTRIBUTIONS Project conception and planning were conducted by D.S. with guidance from R.L. D.S. performed and analysed all experiments except the following:

M.H. participated in the screen, and produced all the data in Supplementary Information, Fig. S2a, b and d except the production and analysis of the extent of the XA12 excision and O.M.

produced Fig. 4c and assisted in staining for Fig. 5h–i. Experimental interpretation was conducted by D.S., M.H. and R.L. The manuscript was written by D.S. and edited by R.L. CORRESPONDING

AUTHOR Correspondence to Ruth Lehmann. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION

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Siekhaus, D., Haesemeyer, M., Moffitt, O. _et al._ RhoL controls invasion and Rap1 localization during immune cell transmigration in _Drosophila_. _Nat Cell Biol_ 12, 605–610 (2010).

https://doi.org/10.1038/ncb2063 Download citation * Received: 25 January 2010 * Accepted: 29 April 2010 * Published: 23 May 2010 * Issue Date: June 2010 * DOI:

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