. 2020 Jul 22;11:842. doi: 10.3389/fphys.2020.00842

TABLE 1.

A toolbox for zebrafish vascular cell biology and selection of recent findings.

Endothelial cell biology applications	Transgenic line name/s	Specific applications for transgenic	Recent biological insights using this strain	References
Cell lineage tracing reporters	TgBAC(etv2:kaede)^ci6	Lineage tracing of angioblasts	Distinct populations of angioblasts give rise to endothelial cells of either the dorsal aorta or the posterior cardinal vein. Angioblasts that originate near the ventral aorta give rise to facial lymphatic endothelial cells. Posterior cardinal vein angioblasts give rise to endothelial cells of the intestinal vessels.	Kohli et al., 2013; Koenig et al., 2016; Eng et al., 2019
	Tg(msgn1:NLS-Kaede)^pc8 Tg(msgn1:Cre-ERT2)^pc9; Tg(actb2:LOXP-AcGFP1-LOXP-mCherry)^pc18	Lineage tracing of somitic origins of endothelial cells	A subset of somite cells (endotome) give rise to angioblasts.	Nguyen et al., 2014
	Tg(EPV.Tp1-Ocu.Hbb2:CreERT2)^jh12; Tg(fli1:LOXP-Cerulean-Hsa.HIST1H2BJ-LOXP,mCherry)^um43	Lineage tracing Notch-active angioblasts	Notch active angioblasts give rise to dorsal aorta endothelial cells but not posterior cardinal vein endothelial cells.	Wang et al., 2011; Quillien et al., 2014
	Tg(EPV.TP1-Mmu.Hbb:Kaede)^um15	Lineage tracing Notch-active endothelial cells	Notch active endothelial cells early in development can give rise to both arterial intersegmental vessel and venous intersegmental vessel endothelial cells. Notch signaling is active specifically in arterial intersegmental vessels even before they anastomose with secondary sprouts to form mature vessels.	Clements et al., 2011; Quillien et al., 2014; Geudens et al., 2019
	Tg(kdrl:NLS-Eos)^ncv6 Tg(Kdrl:Dendra2)^cq52 Tg(fli1a:Gal4FF)^ubs3; Tg(UAS:Kaede)^rk8	Lineage tracing blood vascular endothelial cells	Intersegmental vessel tip cell mitosis results in generation of daughter cells with different size. Later forming angioblasts (25–29 hpf) give rise to endothelial cells in the caudal vessel. Arterial endothelial cells in venous intersegmental vessels migrate dorsally against the flow after secondary sprout anastomosis and are replaced by posterior cardinal vein endothelial cells. Ventral posterior cardinal vein endothelial cells give rise to parachordal lymphatic endothelial cells. Ventral posterior cardinal vein angioblasts give rise to both arterial and venous intestinal vessels.	Hatta et al., 2006; Zygmunt et al., 2011; Fukuhara et al., 2014; Hen et al., 2015; Nicenboim et al., 2015; Costa et al., 2016; Tian et al., 2017; Weijts et al., 2018
	Tg(lyve1b:Kaede)^nz102	Lineage trace ventral aorta angioblasts, and venous/lymphatic endothelial cells	Angioblasts that originate near the ventral aorta give rise to the lymphatic endothelial cells of the facial lymphatics and endothelial cells of the hypobranchial artery.	Eng et al., 2019
Notch signaling reporters	Tg(tp1-MmHbb:EGFP)^um14	Detection of Notch active cells	A subset of angioblasts is Notch active. The tip cell of the venous primordial midbrain channel sprout becomes Notch active prior to fusing with the Notch active arterial system.	Parsons et al., 2009; Quillien et al., 2014; Kaufman et al., 2015; Hasan et al., 2017
	Tg(EPV.Tp1-Mmu.Hbb.d2GFP)^mw43 Tg(EPV.TP1-Mmu.Hbb:Venus-Mmu.Odc1)^s⁹⁴⁰	Dynamic detection of Notch active cells.	Arterial blood flow promotes Notch signaling in arterial intersegmental vessels. Notch signaling in arterial intersegmental vessel prevents it from secondary sprout anastomoses. The tip cell of venous primordial midbrain channel sprout becomes Notch active prior to fusing with the Notch active arterial system.	Clark et al., 2012; Ninov et al., 2012; Hasan et al., 2017; Weijts et al., 2018
	TgBAC(dll4:GAL4FF) ^mu106;Tg(5xUAS:EGFP)^nkuasgfp1a; Tg(EPV.TP1-Mmu.Hbb:hist2h2l-mCherry)^s939	Simultaneous visualization of dll4 transcription and Notch activation	dll4 expression is initiated at the tip cell of venous primordial midbrain channel sprout before it becomes Notch active.	Ninov et al., 2012; Hasan et al., 2017
Ca²⁺ signaling reporters	Tg(10.5xUAS:GCaMP5G)^uq2 Tg(UAS:GCaMP7a)^zf415 Tg(UAS:GCaMP7a)^sh392 Tg(UAS:GCaMP3)^zf350 Tg(Tol2 fliEbasP::mCherry V2A GCaMP6m)^ric100	Visualization of Ca²⁺ signaling	Intersegmental vessel tip cells show Ca²⁺ oscillation that is Vegfa/Kdr/Kdrl-signaling dependent. Intersegmental vessel stalk cells also show Ca²⁺ oscillation. Ca²⁺ oscillation patterns correlate with intersegmental vessel endothelial cell migration and proliferation potential. Calcium signaling increases as dorsal aorta endothelial cells mature. Tmem33 is required for tip endothelial cell Ca²⁺ signaling. The degree of flow-mediated endothelial primary cilia deflection correlates with increased calcium signaling in the dorsal aorta.	Warp et al., 2012; Muto et al., 2013; Goetz et al., 2014; Kita et al., 2015; Yokota et al., 2015; Noren et al., 2016; Lagendijk et al., 2017; Savage et al., 2019
Endothelial cell junction cytoskeleton reporter	Tg(fli1:LIFEACT-EGFP)^zf495 Tg(fli1:LIFEACT-EGFP)^mu240 Tg(fli1:LIFEACT-mClover)^sh467 Tg(UAS:LIFEACT-GFP)^mu271 Tg(4xUAS:Has.UTRN-EGFP)^ubs18	Visualize F-actin polymerization on F-actin-based structures in endothelial cells such as filopodia and endothelial cell junctions	Filopodia are not required for endothelial cell migration but are essential for tip cell anastomosis. Initial contact site of filopodia (junctional spot) and junctional rings on endothelial cells have high F-actin polymerization. F-actin polymerization is required for retraction of blebs during intersegmental vessel lumenization. Dorsal aorta endothelial cells align in the direction of flow as the dorsal aorta matures. Dynamic F-actin polymerization is observed at the Kuglen “neck.” Arterial endothelial cells in venous intersegmental vessels migrate dorsally against the flow after secondary sprout anastomosis and is replaced by posterior cardinal vein endothelial cells. The common cardinal vein lumenize via lumen ensheathment. The lumen and flow are maintained when endothelial cells in multicellular tube divide. Endothelial cells form multicellular tubes using junction-based lamellipodia.	Helker et al., 2013; Phng et al., 2013; Sauteur et al., 2014; Aydogan et al., 2015; Gebala et al., 2016; Hamm et al., 2016; Sauteur et al., 2017; Sugden et al., 2017; Paatero et al., 2018; Weijts et al., 2018; Kugler et al., 2019; Savage et al., 2019
	Tg(4XUAS:mClavGR2-Has.UTRN)^ubs27	Lineage trace endothelial cell junctions	Endothelial cells form multicellular tubes using junction-based lamellipodia.	Paatero et al., 2018
Endothelial cell junctional protein reporter	Tg(14XUAS:EGFP-Hsa.TJP1,myl7:EGFP)^ubs5 Tg(5XUAS:cdh5-EGFP)^ubs12 Tg(fli1:pecam1-EGFP)^ncv27	Visualize ZO1, Ve-cadherin, or Pecam1 localization in endothelial cell junctions.	Junctional spots and junctional rings on endothelial cells have high level of ZO1 and Ve-cadherin. The lumen and flow are maintained when endothelial cells in multicellular tube divide. Endothelial cells form multicellular tubes using junction-based lamellipodia. Intersegmental vessels that remain arterial have a multicellular intersegmental vessel base while intersegmental vessels that will become venous intersegmental vessel have a unicellular base.	Herwig et al., 2011; Lenard et al., 2013; Aydogan et al., 2015; Ando et al., 2016; Paatero et al., 2018; Geudens et al., 2019
Endothelial cell junctional tension sensor	Tg(cdh5:cdh5-TFP-TENS-Venus)^uq11bh	Quantification of Ve-cadherin tension in endothelial cell junctions	Dorsal aorta endothelial cell junctional tension decreases as the dorsal aorta mature.	Lagendijk et al., 2017
Endothelial cell membrane reporter	Tg(kdrl:HsHRAS-mCherry)^s916 Tg(fli1:EGFP-CAAX)^md13 Tg(kdrl:mCherry-CAAX)^y171	Visualize EC membrane and filopodia	Lumenization in the intersegmental vessel is flow-dependent. Cerebral vessels form transient Kuglen structure. Filopodia is not required for endothelial cell migration but is essential for tip cell anastomosis. Initial contact site of filopodia (junctional spot) and junctional rings on endothelial cells have high F-actin polymerization. Lumenization in the intersegmental vessel occur through inverse blebbing. Immature cranial vessels are enriched with primary cilia.	Chi et al., 2008; Fujita et al., 2011; Phng et al., 2013; Gebala et al., 2016; Eisa-Beygi et al., 2018; Kugler et al., 2019
Endothelial cell apical membrane reporter	Tg(fli1:Has.PLCD1-RFP)^md14	Visualize endothelial cell apical membrane	Lumenization in the intersegmental vessel occur through inverse blebbing.	Gebala et al., 2016
Endothelial cell golgi reporter	Tg(fli1:Hsa.B4GALT1-mCherry)^bns9	Visualize endothelial cell polarity	Endothelial cells polarize against flow when blood flow is initiated. Arterial endothelial cells in venous intersegmental vessels migrate dorsally against the flow after secondary sprout anastomosis and is replaced by posterior cardinal vein endothelial cells. The difference in endothelial cell polarity between venous intersegmental vessel and arterial intersegmental vessel endothelial cells is pre-determined prior to secondary sprout anastomosis.	Kwon et al., 2016; Weijts et al., 2018; Geudens et al., 2019
Primary cilia reporter	Tg(actb2:Arl13b-GFP)^hsc5	Visualize primary cilia	Immature cranial vessels are enriched with primary cilia. The degree of flow-mediated endothelial primary cilia deflection correlates with increased calcium signaling in the dorsal aorta.	Borovina et al., 2010; Goetz et al., 2014; Eisa-Beygi et al., 2018
Lymphatic endothelial cell fate reporter	TgBAC(prox1a:KALTA4,4xUAS-E1B:TagRFP)^nim5	Visualize prox1a expression in endothelial cells	prox1a-positive endothelial cells in the posterior cardinal vein undergo mitosis giving rise to a daughter endothelial cell which retains prox1a expression and sprout out of the posterior cardinal vein, and a daughter endothelial cell that lose prox1a expression and remain in the posterior cardinal vein.	Dunworth et al., 2014; Koltowska et al., 2015; Nicenboim et al., 2015
Hyaluronic acid reporter	Tg(ubb:SEC-Rno.Ncan-EGFP)^uq25bh	Visualize hyaluronic acid localization	Hyaluronic acid turnover in the extracellular matrix is essential for proper Vegfa/Kdr/Kdrl signaling during primary angiogenesis.	De Angelis et al., 2017; Grassini et al., 2018
Cell cycle progression reporter	Tg(kdrl:mVenus-gmnn)^ncv3	Visualize endothelial cells in the S/G2/M phase	Intersegmental vessel endothelial cells leaving the dorsal aorta are in the S/G2/M phase and undergo division shortly after.	Fukuhara et al., 2014
Endothelial cell nuclear reporter	Tg(fli1:nEGFP)^y7 Tg(kdrl:nlsmCherry)^is4 Tg(kdrl:nlsEGFP)^ubs1	Visualize EC nucleus	These transgenic lines are widely used to visualize endothelial cell sprouting, migration, division and anastomosis at single cell resolution.	Roman et al., 2002; Blum et al., 2008; Wang et al., 2010