title: CNS angiogenesis and barriergenesis occur simultaneously.
abstract: The blood-brain barrier (BBB) plays a vital role in the central nervous system (CNS). A comprehensive understanding of BBB development has been hampered by difficulties in observing the differentiation of brain endothelial cells (BECs) in real-time. Here, we generated two transgenic zebrafish line, Tg(glut1b:mCherry) and Tg(plvap:EGFP), to serve as in vivo reporters of BBB development. We showed that barriergenesis (i.e. the induction of BEC differentiation) occurs immediately as endothelial tips cells migrate into the brain parenchyma. Using the Tg(glut1b:mCherry) transgenic line, we performed a genetic screen and identified a zebrafish mutant with a nonsense mutation in gpr124, a gene known to play a role in CNS angiogenesis and BBB development. We also showed that our transgenic plvap:EGFP line, a reporter of immature brain endothelium, is initially expressed in newly formed brain endothelial cells, but subsides during BBB maturation. Our results demonstrate the ability to visualize the in vivo differentiation of brain endothelial cells into the BBB phenotype and establish that CNS angiogenesis and barriergenesis occur simultaneously.
authors.affiliation: Department of Chemical Biology & Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA; Integrated Program in Biomedical Sciences, University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Robyn A
authors.initials: RA
authors.lastname: Umans
authors.affiliation: Department of Chemical Biology & Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA; Integrated Program in Biomedical Sciences, University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Hannah E
authors.initials: HE
authors.lastname: Henson
authors.affiliation: Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin, Madison, WI, USA.
authors.forename: Fangzhou
authors.initials: F
authors.lastname: Mu
authors.affiliation: Department of Chemical Biology & Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA.
authors.forename: Chaithanyarani
authors.initials: C
authors.lastname: Parupalli
authors.affiliation: Department of Chemical Biology & Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA.
authors.forename: Bensheng
authors.initials: B
authors.lastname: Ju
authors.affiliation: Cell and Tissue Imaging, St. Jude Children's Research Hospital, Memphis, TN, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Peters
authors.affiliation: Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin, Madison, WI, USA.
authors.forename: Kevin A
authors.initials: KA
authors.lastname: Lanham
authors.affiliation: Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin, Madison, WI, USA.
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Plavicki
authors.affiliation: Department of Chemical Biology & Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA; Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin, Madison, WI, USA. Electronic address: michael.taylor@wisc.edu.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Taylor
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001812:Blood-Brain Barrier / Q000502:physiology*; D002454:Cell Differentiation; D042783:Endothelial Cells / Q000378:metabolism; D017930:Genes, Reporter; D005820:Genetic Testing; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D018919:Neovascularization, Physiologic; D011401:Promoter Regions, Genetic / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D015027:Zebrafish / Q000502:physiology*; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D043562:Receptors, G-Protein-Coupled; D029961:Zebrafish Proteins; C000605631:adgra2 protein, zebrafish; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Angiogenesis; Barriergenesis; Blood-brain barrier (BBB); Brain endothelial cell (BEC); Glucose transporter 1 (Glut1); Plasmalemma vesicle-associated protein (Plvap)