Live-imaging of endothelial Erk activity reveals dynamic and sequential signalling events during regenerative angiogenesis

Kazuhide S Okuda, Mikaela S Keyser, David B Gurevich, Caterina Sturtzel, Elizabeth A Mason, Scott Paterson, Huijun Chen, Mark Scott, Nicholas D Condon, Paul Martin, Martin Distel, Benjamin M Hogan

Research output: Contribution to journalArticlepeer-review

22 Citations (SciVal)


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 Ca 2+ 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.

Original languageEnglish
Article numbere62196
Early online date18 May 2021
Publication statusPublished - 31 May 2021

Bibliographical note

Funding Information:
This work was supported by NHMRC grants 1164734 and 1165117. BMH was supported by an NHMRC fellowship 1155221. We thank Dr. Enid Lam for technical assistance. Imaging was performed in the Australian Cancer Research Foundation’s Cancer Ultrastructure and Function Facility at IMB, Centre for Advanced Histology and Microscopy at Peter MacCallum Cancer Centre, Wolfson Bioimaging Facility at University of Bristol, and the Zebrafish platform Austria for preclinical drug screening at the Children’s Cancer Research Institute supported by the Austrian Research Promotion Agency (FFG) project 7640628 (Danio4Can). We thank Olympus for use of the Olympus Yokogawa CSU-W1 Spinning Disc Confocal microscope.

Data availability
All data generated or analysed during this study are included in the manuscript and supporting files.

ASJC Scopus subject areas

  • General Neuroscience
  • General Biochemistry,Genetics and Molecular Biology
  • General Immunology and Microbiology


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