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Vegfr3-CreER T2 mouse, a new genetic tool for targeting the lymphatic system

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An Erratum to this article was published on 06 June 2016

Abstract

The lymphatic system is essential in many physiological and pathological processes. Still, much remains to be known about the molecular mechanisms that control its development and function and how to modulate them therapeutically. The study of these mechanisms will benefit from better controlled genetic mouse models targeting specifically lymphatic endothelial cells. Among the genes expressed predominantly in lymphatic endothelium, Vegfr3 was the first one identified and is still considered to be one of the best lymphatic markers and a key regulator of the lymphatic system. Here, we report the generation of a Vegfr3-CreER T2 knockin mouse by gene targeting in embryonic stem cells. This mouse expresses the tamoxifen-inducible CreERT2 recombinase under the endogenous transcriptional control of the Vegfr3 gene without altering its physiological expression or regulation. The Vegfr3-CreER T2 allele drives efficient recombination of floxed sequences upon tamoxifen administration specifically in Vegfr3-expressing cells, both in vitro, in primary lymphatic endothelial cells, and in vivo, at different stages of mouse embryonic development and postnatal life. Thus, our Vegfr3-CreER T2 mouse constitutes a new powerful genetic tool for lineage tracing analysis and for conditional gene manipulation in the lymphatic endothelium that will contribute to improve our current understanding of this system.

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Acknowledgments

The support of the Spanish National Cancer Research Centre (CNIO) Transgenesis, Comparative Pathology and Animal Facility Core Units is greatly acknowledged. We also thank the Uppsala University Animal Units for animal husbandry. This work was supported by the Ministry of Science and Innovation of Spain: Grants BIO2009-09488 and BIO2006-03213 (to S.O.) and FPU fellowship (to IM-C), the CNIO, the Lymphatic Education and Research Network (postdoctoral fellowship to M.F.) and the Swedish Research Council and European Research Council (ERC-2014-CoG 646849) (to T.M.).

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Author notes

  1. Lukas Stanczuk

    Present address: Cambridge Cancer Centre, Cambridge, UK

  2. Rodrigo Diéguez-Hurtado

    Present address: Department of Tissue Morphogenesis, Max Planck Institute for Molecular Biomedicine, Münster, Germany

  3. David Olmeda

    Present address: Melanoma Group, Molecular Oncology Programme, Spanish National Cancer Research Centre, Madrid, Spain

Authors and Affiliations

  1. Transgenic Mice Unit, Biotechnology Programme, Spanish National Cancer Research Centre, Madrid, Spain

    Ines Martinez-Corral, Rodrigo Diéguez-Hurtado, David Olmeda & Sagrario Ortega

  2. Department of Immunology, Genetics and Pathology, Uppsala University, 752 85, Uppsala, Sweden

    Ines Martinez-Corral, Lukas Stanczuk, Maike Frye, Maria Helena Ulvmar & Taija Makinen

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Correspondence to Sagrario Ortega.

Electronic supplementary material

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10456_2016_9505_MOESM1_ESM.tif

Supplementary Figure 1: Effect of prolonged TAM diet in Vegfr3-CreER T2 ; R26-LSL-LacZ heterozygous mice. a) Whole mount (diaphragm and peritoneum) or frozen sections from different organs from Vegfr3-CreER T2 ; R26-LSL-lacZ mice, stained with X-Gal for LacZ expression. Mice were treated during 3, 5 or 7 consecutive weeks with TAM diet, starting at 4 weeks of age. Scale bars: 100 μm (Frozen sections), 1 mm (whole mount). (TIFF 6483 kb)

Supplementary material 2 (DOCX 17 kb)

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Martinez-Corral, I., Stanczuk, L., Frye, M. et al. Vegfr3-CreER T2 mouse, a new genetic tool for targeting the lymphatic system. Angiogenesis 19, 433–445 (2016). https://doi.org/10.1007/s10456-016-9505-x

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