Endothelial Ca2+ oscillations reflect VEGFR signaling-regulated angiogenic capacity in vivo

Abstract
Article and author information
Metrics

Abstract

Sprouting angiogenesis is a well-coordinated process controlled by multiple extracellular inputs, including vascular endothelial growth factor (VEGF). However, little is known about when and how individual endothelial cell (EC) responds to angiogenic inputs in vivo. Here, we visualized endothelial Ca²⁺ dynamics in zebrafish and found that intracellular Ca²⁺ oscillations occurred in ECs exhibiting angiogenic behavior. Ca²⁺ oscillations depended upon Vegfr2 and Vegfr3 in ECs budding from the dorsal aorta (DA) and posterior cardinal vein, respectively. Thus, visualizing Ca²⁺ oscillations allowed us to monitor EC responses to angiogenic cues. Vegfr-dependent Ca²⁺ oscillations occurred in migrating tip cells as well as stalk cells budding from the DA. We investigated how Dll4/Notch signaling regulates endothelial Ca²⁺ oscillations and found that it was required for the selection of single stalk cell as well as tip cell. Thus, we captured spatio-temporal Ca²⁺ dynamics during sprouting angiogenesis, as a result of cellular responses to angiogenic inputs.

Article and author information

Author details

Yasuhiro Yokota

Department of Cell Biology, National Cerebral and Cardiovascular Cener, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Hiroyuki Nakajima

Department of Cell Biology, National Cerebral and Cardiovascular Center, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Yuki Wakayama

Department of Cell Biology, National Cerebral and Cardiovascular Center, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Akira Muto

Division of Molecular and Developmental Biology, National Institute of Genetics, Department of Genetics, Graduate University for Advanced Studies, Mishima, Japan

Competing interests
The authors declare that no competing interests exist.
Koichi Kawakami

Division of Molecular and Developmental Biology, National Institute of Genetics, Department of Genetics, National Institute of Genetics,, Mishima, Japan

Competing interests
The authors declare that no competing interests exist.
Shigetomo Fukuhara

Department of Cell Biology, National Cerebral and Cardiovascular Center, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Naoki Mochizuki

Department of Cell Biology, National Cerebral and Cardiovascular Center, Suita, Japan

For correspondence
nmochizu@ri.ncvc.go.jp

Competing interests
The authors declare that no competing interests exist.

Ethics

Animal experimentation: Animal experimentation: The experiments using zebrafish were approved by the animal committee of National Cerebral and Cardiovascular Center (No. 14005) and performed according to the guidance of the Institute.

Copyright

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.