Live-imaging of endothelial Erk activity reveals dynamic and sequential signalling events during regenerative angiogenesis
- Peter MacCallum Cancer Centre, Australia
- University of Queensland, Australia
- University of Bristol, United Kingdom
- St. Anna Kinderkrebsforschung, Children's Cancer Research Institute, Austria
Abstract
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. 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 concomitant 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.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files.
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Funding
National Health and Medical Research Council (1164734)
- Benjamin M Hogan
National Health and Medical Research Council (1165117)
- Benjamin M Hogan
Austrian Research Promotion Agency (7640628)
- Martin Distel
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All zebrafish work was conducted in accordance with the guidelines of the animal ethics committees at the University of Queensland (AE54297), University of Melbourne, Peter MacCallum Cancer Centre (E634 and E643), University of Bristol (3003318), and the Children's Cancer Research Institute (GZ:565304/2014/6 and GZ:534619/2014/4).
Copyright
© 2021, Okuda et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Live-imaging of endothelial Erk activity reveals dynamic and sequential signalling events during regenerative angiogenesis
eLife 10:e62196.
https://doi.org/10.7554/eLife.62196
