Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia; Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia; Department of Anatomy and Physiology, University of Melbourne, Melbourne, Australia; Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
Mikaela S Keyser
Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
David B Gurevich
School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol, Bristol, United Kingdom; Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, United Kingdom
Caterina Sturtzel
Innovative Cancer Models, St Anna Kinderkrebsforschung, Children’s Cancer Research Institute, Vienna, Austria; Zebrafish Platform Austria for preclinical drug screening (ZANDR), Vienna, Austria
Elizabeth A Mason
Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia; Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia; Department of Anatomy and Physiology, University of Melbourne, Melbourne, Australia
Scott Paterson
Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia; Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia; Department of Anatomy and Physiology, University of Melbourne, Melbourne, Australia; Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
Huijun Chen
Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
Mark Scott
Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
Nicholas D Condon
Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
Paul Martin
School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol, Bristol, United Kingdom
Innovative Cancer Models, St Anna Kinderkrebsforschung, Children’s Cancer Research Institute, Vienna, Austria; Zebrafish Platform Austria for preclinical drug screening (ZANDR), Vienna, Austria
Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia; Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia; Department of Anatomy and Physiology, University of Melbourne, Melbourne, Australia; Institute for Molecular Bioscience, The University of Queensland, St Lucia, St Lucia, Australia
The formation of new blood vessel networks occurs via angiogenesis during development, tissue repair, and disease. Angiogenesis is regulated by intracellular endothelial signalling pathways, induced downstream of vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs). A major challenge in understanding angiogenesis is interpreting how signalling events occur dynamically within endothelial cell populations during sprouting, proliferation, and migration. Extracellular signal-regulated kinase (Erk) is a central downstream effector of Vegf-signalling and reports the signalling that drives angiogenesis. We generated a vascular Erk biosensor transgenic line in zebrafish using a kinase translocation reporter that allows live-imaging of Erk-signalling dynamics. We demonstrate the utility of this line to live-image Erk activity during physiologically relevant angiogenic events. Further, we reveal dynamic and sequential endothelial cell Erk-signalling events following blood vessel wounding. Initial signalling is dependent upon Ca2+ in the earliest responding endothelial cells, but is independent of Vegfr-signalling and local inflammation. The sustained regenerative response, however, involves a Vegfr-dependent mechanism that initiates concomitantly with the wound inflammatory response. This work reveals a highly dynamic sequence of signalling events in regenerative angiogenesis and validates a new resource for the study of vascular Erk-signalling in real-time.
