Vegfr3-tdTomato, a reporter mouse for microscopic visualization of lymphatic vessel by multiple modalities

PLoS One. 2021 Sep 20;16(9):e0249256. doi: 10.1371/journal.pone.0249256. eCollection 2021.

Abstract

Lymphatic vessels are indispensable for tissue fluid homeostasis, transport of solutes and dietary lipids and immune cell trafficking. In contrast to blood vessels, which are easily visible by their erythrocyte cargo, lymphatic vessels are not readily detected in the tissue context. Their invisibility interferes with the analysis of the three-dimensional lymph vessel structure in large tissue volumes and hampers dynamic intravital studies on lymphatic function and pathofunction. An approach to overcome these limitations are mouse models, which express transgenic fluorescent proteins under the control of tissue-specific promotor elements. We introduce here the BAC-transgenic mouse reporter strain Vegfr3-tdTomato that expresses a membrane-tagged version of tdTomato under control of Flt4 regulatory elements. Vegfr3-tdTomato mice inherited the reporter in a mendelian fashion and showed selective and stable fluorescence in the lymphatic vessels of multiple organs tested, including lung, kidney, heart, diaphragm, intestine, mesentery, liver and dermis. In this model, tdTomato expression was sufficient for direct visualisation of lymphatic vessels by epifluorescence microscopy. Furthermore, lymph vessels were readily visualized using a number of microscopic modalities including confocal laser scanning, light sheet fluorescence and two-photon microscopy. Due to the early onset of VEGFR-3 expression in venous embryonic vessels and the short maturation time of tdTomato, this reporter offers an interesting alternative to Prox1-promoter driven lymphatic reporter mice for instance to study the developmental differentiation of venous to lymphatic endothelial cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Endothelial Cells
  • Genes, Reporter
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Lymphatic Vessels / cytology*
  • Lymphatic Vessels / physiology
  • Mice, Transgenic*
  • Microscopy, Confocal
  • Microscopy, Fluorescence / methods
  • Red Fluorescent Protein
  • Vascular Endothelial Growth Factor Receptor-3 / genetics*
  • Vascular Endothelial Growth Factor Receptor-3 / metabolism

Substances

  • Luminescent Proteins
  • Vascular Endothelial Growth Factor Receptor-3

Associated data

  • figshare/10.6084/m9.figshare.14931708.v1

Grants and funding

FK and ER are funded by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - SFB1348/1 – 386797833 (https://gepris.dfg.de/gepris/projekt/386797833) and FK by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - SFB1450/1 - 431460824 (https://gepris.dfg.de/gepris/projekt/431460824). FK and RH are funded by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - SFB656/3 – 12467772 (https://gepris.dfg.de/gepris/projekt/12467772). NK is funded by CiM-IMPRS, the joint graduate school of the Cells-in-Motion Cluster of Excellence (EXC 1003 - CiM), University of Münster, Germany and the International Max Planck Research School - Molecular Biomedicine, Münster, Germany (https://gepris.dfg.de/gepris/projekt/194347757). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.