Src- and Fyn-dependent apical membrane trafficking events control endothelial lumen formation during vascular tube morphogenesis

Dae Joong Kim, Pieter R. Norden, Jocelynda Salvador, David M. Barry, Stephanie L.K. Bowers, Ondine Cleaver, George E. Davis

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Here we examine the question of how endothelial cells (ECs) develop their apical membrane surface domain during lumen and tube formation. We demonstrate marked apical membrane targeting of activated Src kinases to this apical domain during early and late stages of this process. Immunostaining for phosphotyrosine or phospho-Src reveals apical membrane staining in intracellular vacuoles initially. This is then followed by vacuole to vacuole fusion events to generate an apical luminal membrane, which is similarly decorated with activated phospho-Src kinases. Functional blockade of Src kinases completely blocks EC lumen and tube formation, whether this occurs during vasculogenic tube assembly or angiogenic sprouting events. Multiple Src kinases participate in this apical membrane formation process and siRNA suppression of Src, Fyn and Yes, but not Lyn, blocks EC lumen formation. We also demonstrate strong apical targeting of Src-GFP and Fyn-GFP fusion proteins and increasing their expression enhances lumen formation. Finally, we show that Src- and Fyn-associated vacuoles track and fuse along a subapically polarized microtubule cytoskeleton, which is highly acetylated. These vacuoles generate the apical luminal membrane in a stereotypically polarized, perinuclear position. Overall, our study identifies a critical role for Src kinases in creating and decorating the EC apical membrane surface during early and late stages of lumen and tube formation, a central event in the molecular control of vascular morphogenesis.

Original languageEnglish (US)
Article numbere0184461
JournalPLoS One
Volume12
Issue number9
DOIs
StatePublished - Sep 1 2017

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Fingerprint Dive into the research topics of 'Src- and Fyn-dependent apical membrane trafficking events control endothelial lumen formation during vascular tube morphogenesis'. Together they form a unique fingerprint.

  • Cite this