Generating Vegfr3 reporter transgenic mouse expressing membrane-tagged Venus for visualization of VEGFR3 expression in vascular and lymphatic endothelial cells.

Chisato Watanabe,Satoru Takahashi,Takuya Azami,Jun Matsushita,Tomoyuki Tsukiyama,Masatsugu Ema,Seiya Mizuno,Setsuko Tsukiyama-Fujii,Masuko Ushio-Fukai

doi:10.1371/journal.pone.0210060

Chisato Watanabe, Satoru Takahashi + Show 7 more

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https://doi.org/10.1371/journal.pone.0210060

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Abstract

Vascular endothelial growth factor receptor 3 (Vegfr3) has been widely used as a marker for lymphatic and vascular endothelial cells during mouse embryonic development and in adult mouse, making it valuable for studying angiogenesis and lymphangiogenesis under normal and pathological conditions. Here, we report the generation of a novel transgenic (Tg) mouse that expresses a membrane-localized fluorescent reporter protein, Gap43-Venus, under the control of the Vegfr3 regulatory sequence. Vegfr3-Gap43-Venus BAC Tg recapitulated endogenous Vegfr3 expression in vascular and lymphatic endothelial cells during embryonic development and tumor development. Thus, this Tg mouse line contributes a valuable model to study angiogenesis and lymphangiogenesis in physiological and pathological contexts.

Highlights

The vascular endothelial growth factors (VEGFs) play as critical regulators of vascular development among mammalian species [1]
Because VEGFR3 expression has been reported to be downregulated in vascular endothelial cells at the dorsal aorta, which restricts to lymphatic endothelial cells in the skin around E14.5 [5], we examined Venus expression in the back skin of E14.5 embryos and found that Venus expression was overlapped with that of endogenous VEGFR3 and Lyve1-positive lymphatic endothelial cells (Fig 4A)
We demonstrated that Vascular endothelial growth factor receptor 3 (Vegfr3)-Gap43-Venus BAC Tg can recapitulate endogenous VEGFR3 expression in vascular endothelial cells and lymphatic endothelial cells during embryonic development and in adulthood

Summary

Introduction

The vascular endothelial growth factors (VEGFs) play as critical regulators of vascular development among mammalian species [1]. The VEGF family consists of six secreted proteins (VEGF-A, B, C, D, E and placental growth factor), which have different binding affinities for three tyrosine kinase receptors; VEGFR1 (Fms-like tyrosine kinase 1; Flt1), VEGFR2 (Fetal liver kinase 1; Flk1), VEGFR3 (Fms-like tyrosine kinase 4; Flt4)[1,2]. VEGF family members bind to non-tyrosine kinase receptors, Neuropilin and 2, which are considered to function as co-receptors for the VEGFRs. VEGF-A and VEGF-B and placental growth factor bind to VEGFR1, VEGF-A and VEGF-C to VEGFR2, and VEGF-C and VEGF-D to VEGFR3. VEGF-C binds to VEGFR2/VEGFR3 heterodimer and transduces signals through Akt, whereas VEGFR3/VEGFR3 homodimer transduces signals through ERK [3].

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