A transgenic Prox1-Cre-tdTomato reporter mouse for lymphatic vessel research

PLoS One. 2015 Apr 7;10(4):e0122976. doi: 10.1371/journal.pone.0122976. eCollection 2015.

Abstract

The lymphatic vascular system plays an active role in immune cell trafficking, inflammation and cancer spread. In order to provide an in vivo tool to improve our understanding of lymphatic vessel function in physiological and pathological conditions, we generated and characterized a tdTomato reporter mouse and crossed it with a mouse line expressing Cre recombinase under the control of the lymphatic specific promoter Prox1 in an inducible fashion. We found that the tdTomato fluorescent signal recapitulates the expression pattern of Prox1 in lymphatic vessels and other known Prox1-expressing organs. Importantly, tdTomato co-localized with the lymphatic markers Prox1, LYVE-1 and podoplanin as assessed by whole-mount immunofluorescence and FACS analysis. The tdTomato reporter was brighter than a previously established red fluorescent reporter line. We confirmed the applicability of this animal model to intravital microscopy of dendritic cell migration into and within lymphatic vessels, and to fluorescence-activated single cell analysis of lymphatic endothelial cells. Additionally, we were able to describe the early morphological changes of the lymphatic vasculature upon induction of skin inflammation. The Prox1-Cre-tdTomato reporter mouse thus shows great potential for lymphatic research.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Movement
  • Cell Tracking
  • Dendritic Cells / physiology
  • Dermatitis / physiopathology
  • Gene Expression
  • Genes, Reporter
  • HEK293 Cells
  • Humans
  • Integrases / genetics
  • Luminescent Proteins / biosynthesis*
  • Luminescent Proteins / genetics
  • Lymphatic Vessels / physiopathology*
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Organ Specificity
  • Red Fluorescent Protein
  • Transgenes

Substances

  • Luminescent Proteins
  • Cre recombinase
  • Integrases

Grants and funding

This work was supported by Swiss National Science Foundation grant 31003A-130627, Advanced European Research Council Grant LYVICAM, the Leducq Foundation Transatlantic Network of Excellence grant Lymph Vessels in Obesity and Cardiovascular Disease, Oncosuisse and Krebsliga Zurich (to M.D.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.