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Research Article
Olfactomedin-1 activity identifies a cell invasion checkpoint during epithelial-mesenchymal transition in the chick embryonic heart
Alejandro Lencinas, Danny C. Chhun, Kelvin P. Dan, Kristen D. Ross, Elizabeth A. Hoover, Parker B. Antin, Raymond B. Runyan
Disease Models & Mechanisms 2013 : dmm.010595 doi: 10.1242/dmm.010595 Published 8 February 2013
Alejandro Lencinas
University of Arizona, Tucson, Arizona, USA
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Danny C. Chhun
University of Arizona, Tucson, Arizona, USA
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Kelvin P. Dan
University of Arizona, Tucson, Arizona, USA
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Kristen D. Ross
University of Arizona, Tucson, Arizona, USA
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Elizabeth A. Hoover
University of Arizona, Tucson, Arizona, USA
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Parker B. Antin
University of Arizona, Tucson, Arizona, USA
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Raymond B. Runyan
University of Arizona, Tucson, Arizona, USA
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Summary

Endothelia in the atrioventricular (AV) canal of the developing heart undergo a prototypical epithelial mesenchymal transition (EMT) to begin heart valve formation. Using an in vitro invasion assay, an extracellular matrix protein, Olfactomedin-1 (OLFM1), was found to increase mesenchymal cell numbers in AV canals from embryonic chick hearts. Treatment with both anti-OLFM1 antibody and siRNA targeting OLFM1 inhibits mesenchymal cell formation. OLFM1 does not alter cell proliferation, migration or apoptosis. Dispersion, but lack of invasion in the presence of inhibiting antibody, identifies a specific role for OLFM1 in cell invasion during EMT. This role is conserved in other epithelia, as OLFM1 similarly enhances invasion by MDCK epithelial cells in a transwell assay. Synergy is observed when TGFβ2 and OLFM1 are added to MDCK cell cultures, indicating that OLFM-1 activity is cooperative with TGFβ. Inhibition of both OLFM1 and TGFβ in heart invasion assays shows a similar cooperative role during development. To explore OLFM1 activity during EMT, representative EMT markers were examined. Effects of OLFM1 protein and anti-OLFM1 on transcripts of cell-cell adhesion molecules and the transcription factors Snail-1, Snail-2, Twist1 and Sox-9 argue that OLFM1 does not initiate EMT. Rather, regulation of transcripts of Zeb1 and Zeb2, secreted proteases and mesenchymal cell markers by both OLFM1 and anti-OLFM1 is consistent with regulation of the cell invasion step of EMT. We conclude that OLFM1 is present and necessary during EMT in the embryonic chick heart. Its role in cell invasion and mesenchymal cell gene regulation suggests an invasion checkpoint in EMT where OLFM1 acts to promote cell invasion into the three-dimensional matrix.

  • Received July 20, 2012.
  • Accepted December 12, 2012.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0), which permits unrestricted non-commercial use, distribution and reproduction in any medium provided that the original work is properly cited and all further distributions of the work or adaptation are subject to the same Creative Commons License terms.

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Olfactomedin-1 activity identifies a cell invasion checkpoint during epithelial-mesenchymal transition in the chick embryonic heart
Alejandro Lencinas, Danny C. Chhun, Kelvin P. Dan, Kristen D. Ross, Elizabeth A. Hoover, Parker B. Antin, Raymond B. Runyan
Disease Models & Mechanisms 2013 : dmm.010595 doi: 10.1242/dmm.010595 Published 8 February 2013
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Olfactomedin-1 activity identifies a cell invasion checkpoint during epithelial-mesenchymal transition in the chick embryonic heart
Alejandro Lencinas, Danny C. Chhun, Kelvin P. Dan, Kristen D. Ross, Elizabeth A. Hoover, Parker B. Antin, Raymond B. Runyan
Disease Models & Mechanisms 2013 : dmm.010595 doi: 10.1242/dmm.010595 Published 8 February 2013

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