Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo

Janna Krueger, Dong Liu, Katja Scholz, Anja Zimmer, Yu Shi, Christian Klein, Arndt Siekmann, Stefan Schulte-Merker, Melissa Cudmore, Asif Ahmed, Ferdinand le Noble

Research output: Contribution to journalArticle

Abstract

Endothelial tip cells guide angiogenic sprouts by exploring the local environment for guidance cues such as vascular endothelial growth factor (VegfA). Here we present Flt1 (Vegf receptor 1) loss- and gain-of-function data in zebrafish showing that Flt1 regulates tip cell formation and arterial branching morphogenesis. Zebrafish embryos expressed soluble Flt1 (sFlt1) and membrane-bound Flt1 (mFlt1). In Tg(flt1(BAC):yfp) × Tg(kdrl:ras-cherry)(s916) embryos, flt1:yfp was expressed in tip, stalk and base cells of segmental artery sprouts and overlapped with kdrl:cherry expression in these domains. flt1 morphants showed increased tip cell numbers, enhanced angiogenic behavior and hyperbranching of segmental artery sprouts. The additional arterial branches developed into functional vessels carrying blood flow. In support of a functional role for the extracellular VEGF-binding domain of Flt1, overexpression of sflt1 or mflt1 rescued aberrant branching in flt1 morphants, and overexpression of sflt1 or mflt1 in controls resulted in short arterial sprouts with reduced numbers of filopodia. flt1 morphants showed reduced expression of Notch receptors and of the Notch downstream target efnb2a, and ectopic expression of flt4 in arteries, consistent with loss of Notch signaling. Conditional overexpression of the notch1a intracellular cleaved domain in flt1 morphants restored segmental artery patterning. The developing nervous system of the trunk contributed to the distribution of Flt1, and the loss of flt1 affected neurons. Thus, Flt1 acts in a Notch-dependent manner as a negative regulator of tip cell differentiation and branching. Flt1 distribution may be fine-tuned, involving interactions with the developing nervous system.
Original languageEnglish
Pages (from-to)2111-2120
Number of pages10
JournalDevelopment
Volume138
Issue number10
DOIs
Publication statusPublished - 15 May 2011

Fingerprint

Zebrafish
Morphogenesis
Embryonic Structures
Arteries
Vascular Endothelial Growth Factor A
Nervous System
Notch Receptors
Pseudopodia
Cues
Blood Vessels
Cell Differentiation
Endothelial Cells
Cell Count
Neurons
Membranes

Keywords

  • Flt1
  • VEGF
  • angiogenesis
  • Notch
  • tip cells
  • nerves
  • zebrafish

Cite this

Krueger, J., Liu, D., Scholz, K., Zimmer, A., Shi, Y., Klein, C., ... le Noble, F. (2011). Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo. Development, 138(10), 2111-2120. https://doi.org/10.1242/dev.063933
Krueger, Janna ; Liu, Dong ; Scholz, Katja ; Zimmer, Anja ; Shi, Yu ; Klein, Christian ; Siekmann, Arndt ; Schulte-Merker, Stefan ; Cudmore, Melissa ; Ahmed, Asif ; le Noble, Ferdinand. / Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo. In: Development. 2011 ; Vol. 138, No. 10. pp. 2111-2120.
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Krueger, J, Liu, D, Scholz, K, Zimmer, A, Shi, Y, Klein, C, Siekmann, A, Schulte-Merker, S, Cudmore, M, Ahmed, A & le Noble, F 2011, 'Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo', Development, vol. 138, no. 10, pp. 2111-2120. https://doi.org/10.1242/dev.063933

Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo. / Krueger, Janna; Liu, Dong; Scholz, Katja; Zimmer, Anja; Shi, Yu; Klein, Christian; Siekmann, Arndt; Schulte-Merker, Stefan; Cudmore, Melissa; Ahmed, Asif; le Noble, Ferdinand.

In: Development, Vol. 138, No. 10, 15.05.2011, p. 2111-2120.

Research output: Contribution to journalArticle

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T1 - Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo

AU - Krueger, Janna

AU - Liu, Dong

AU - Scholz, Katja

AU - Zimmer, Anja

AU - Shi, Yu

AU - Klein, Christian

AU - Siekmann, Arndt

AU - Schulte-Merker, Stefan

AU - Cudmore, Melissa

AU - Ahmed, Asif

AU - le Noble, Ferdinand

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AB - Endothelial tip cells guide angiogenic sprouts by exploring the local environment for guidance cues such as vascular endothelial growth factor (VegfA). Here we present Flt1 (Vegf receptor 1) loss- and gain-of-function data in zebrafish showing that Flt1 regulates tip cell formation and arterial branching morphogenesis. Zebrafish embryos expressed soluble Flt1 (sFlt1) and membrane-bound Flt1 (mFlt1). In Tg(flt1(BAC):yfp) × Tg(kdrl:ras-cherry)(s916) embryos, flt1:yfp was expressed in tip, stalk and base cells of segmental artery sprouts and overlapped with kdrl:cherry expression in these domains. flt1 morphants showed increased tip cell numbers, enhanced angiogenic behavior and hyperbranching of segmental artery sprouts. The additional arterial branches developed into functional vessels carrying blood flow. In support of a functional role for the extracellular VEGF-binding domain of Flt1, overexpression of sflt1 or mflt1 rescued aberrant branching in flt1 morphants, and overexpression of sflt1 or mflt1 in controls resulted in short arterial sprouts with reduced numbers of filopodia. flt1 morphants showed reduced expression of Notch receptors and of the Notch downstream target efnb2a, and ectopic expression of flt4 in arteries, consistent with loss of Notch signaling. Conditional overexpression of the notch1a intracellular cleaved domain in flt1 morphants restored segmental artery patterning. The developing nervous system of the trunk contributed to the distribution of Flt1, and the loss of flt1 affected neurons. Thus, Flt1 acts in a Notch-dependent manner as a negative regulator of tip cell differentiation and branching. Flt1 distribution may be fine-tuned, involving interactions with the developing nervous system.

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KW - VEGF

KW - angiogenesis

KW - Notch

KW - tip cells

KW - nerves

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