RRC ID 49281
Author Fukuhara S, Zhang J, Yuge S, Ando K, Wakayama Y, Sakaue-Sawano A, Miyawaki A, Mochizuki N.
Title Visualizing the cell-cycle progression of endothelial cells in zebrafish.
Journal Dev Biol
Abstract The formation of vascular structures requires precisely controlled proliferation of endothelial cells (ECs), which occurs through strict regulation of the cell cycle. However, the mechanism by which EC proliferation is coordinated during vascular formation remains largely unknown, since a method of analyzing cell-cycle progression of ECs in living animals has been lacking. Thus, we devised a novel system allowing the cell-cycle progression of ECs to be visualized in vivo. To achieve this aim, we generated a transgenic zebrafish line that expresses zFucci (zebrafish fluorescent ubiquitination-based cell cycle indicator) specifically in ECs (an EC-zFucci Tg line). We first assessed whether this system works by labeling the S phase ECs with EdU, then performing time-lapse imaging analyses and, finally, examining the effects of cell-cycle inhibitors. Employing the EC-zFucci Tg line, we analyzed the cell-cycle progression of ECs during vascular development in different regions and at different time points and found that ECs proliferate actively in the developing vasculature. The proliferation of ECs also contributes to the elongation of newly formed blood vessels. While ECs divide during elongation in intersegmental vessels, ECs proliferate in the primordial hindbrain channel to serve as an EC reservoir and migrate into basilar and central arteries, thereby contributing to new blood vessel formation. Furthermore, while EC proliferation is not essential for the formation of the basic framework structures of intersegmental and caudal vessels, it appears to be required for full maturation of these vessels. In addition, venous ECs mainly proliferate in the late stage of vascular development, whereas arterial ECs become quiescent at this stage. Thus, we anticipate that the EC-zFucci Tg line can serve as a tool for detailed studies of the proliferation of ECs in various forms of vascular development in vivo.
Volume 393(1)
Pages 10-23
Published 2014-9-1
DOI 10.1016/j.ydbio.2014.06.015
PII S0012-1606(14)00311-X
PMID 24975012
MeSH Animals Animals, Genetically Modified / embryology Animals, Genetically Modified / genetics Cell Cycle / genetics* Cell Cycle / physiology Cell Division Cell Proliferation Cyclin-Dependent Kinase Inhibitor p27 / genetics Endothelium, Vascular / cytology Endothelium, Vascular / physiology* Gene Knockdown Techniques Intracellular Signaling Peptides and Proteins / genetics Membrane Proteins / genetics Morpholinos / genetics Neovascularization, Physiologic / genetics* Zebrafish / genetics Zebrafish / physiology* Zebrafish Proteins / biosynthesis
IF 2.896
Times Cited 29
DNA material mCherry-zCdt1(1/190) / pT2KXIGdeltain (RDB15278) mVenus-zGeminin (1/100) / pT2KXIGdeltain (RDB15279).