We have applied search algorithms to expression databases to identify genes whose expression is restricted to the endothelial cell. Such genes frequently play a critical role in endothelial biology and angiogenesis. Two such genes are the roundabout receptor Robo4 and the ECSCR (endothelial-cell-specific chemotaxis regulator). Endothelial cells express both Robo1 and Robo4, which we have knocked down using siRNA (small interfering RNA) and then studied the effect in a variety of in vitro assays. Both Robo4 and Robo1 knockdown inhibited in vitro tube formation on Matrigel™. Transfection of Robo4 into endothelial cells increased the number of filopodial extensions from the cell, but failed to do so in Robo1-knockdown cells. Separate immunoprecipitation studies showed that Robo1 and Robo4 heterodimerize. We conclude from this and other work that a heteroduplex of Robo1 and Robo4 signals through WASP (Wiskott–Aldrich syndrome protein) and other actin nucleation-promoting factors to increase the number of filopodia and cell migration. Knockdown of the transmembrane ECSCR protein in endothelial cells also reduced chemotaxis and impaired tube formation on Matrigel™. Yeast two-hybrid analysis and immunoprecipitation studies showed that, in contrast with the roundabouts, ECSCR binds to the actin-modulatory filamin A. We conclude that all three of these genes are critical for effective endothelial cell migration and, in turn, angiogenesis.
Skip Nav Destination
Article navigation
December 2009
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
Conference Article|
November 19 2009
Functionally defining the endothelial transcriptome, from Robo4 to ECSCR
Ana Raquel Verissimo;
Ana Raquel Verissimo
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
John M.J. Herbert;
John M.J. Herbert
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
Victoria L. Heath;
Victoria L. Heath
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
John A. Legg;
John A. Legg
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
Helen Sheldon;
Helen Sheldon
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
Maud Andre;
Maud Andre
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
Rajeeb Kumar Swain;
Rajeeb Kumar Swain
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
Search for other works by this author on:
Roy Bicknell
Roy Bicknell
1
1Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K.
1To whom correspondence should be addressed (email: r.bicknell@bham.ac.uk).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
July 06 2009
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© The Authors Journal compilation © 2009 Biochemical Society
2009
Biochem Soc Trans (2009) 37 (6): 1214–1217.
Article history
Received:
July 06 2009
Citation
Ana Raquel Verissimo, John M.J. Herbert, Victoria L. Heath, John A. Legg, Helen Sheldon, Maud Andre, Rajeeb Kumar Swain, Roy Bicknell; Functionally defining the endothelial transcriptome, from Robo4 to ECSCR. Biochem Soc Trans 1 December 2009; 37 (6): 1214–1217. doi: https://doi.org/10.1042/BST0371214
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.