April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Cellular mechanisms of corneal lymphangiogenesis revealed by live imaging
Author Affiliations & Notes
  • Richard M Tempero
    Boys Town National Rsch Hosp, Omaha, NE
  • Alicia L Connor
    Boys Town National Rsch Hosp, Omaha, NE
  • Philip M Kelley
    Boys Town National Rsch Hosp, Omaha, NE
  • Footnotes
    Commercial Relationships Richard Tempero, None; Alicia Connor, None; Philip Kelley, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 5047. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Richard M Tempero, Alicia L Connor, Philip M Kelley; Cellular mechanisms of corneal lymphangiogenesis revealed by live imaging. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5047.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: The lymphatic vasculature regulates normal physiology and plays an important role in corneal inflammatory disease. Here we investigate the cellular mechanisms and proliferative capacities of newly synthesized lymphatic vessels using inductive genetic lineage tracing studies and quantitative analysis.

Methods: Transgenic mice were developed that carry an inducible CRE recombinase-estrogen receptor construct targeted to lymphatic endothelial cells (LECs) and a td tomato reporter gene. We used this inductive transgenic strategy in adult mice to label a sample population of LECs. We followed the fate of their progeny at single cell resolution in real time using intravital microscopy during suture induced corneal lymphangiogenesis. We performed morphologic and quantitative analysis to describe mechanisms of growth.

Results: Using high dose inductive conditions we visualized and quantified a dynamic process of lymphatic vessel growth and contraction, and diverse proliferative capacities in td tomato+ LECs during lymphangiogenesis. The majority of the new growth was generated by sprouting from the preexisting limbal lymphatic vessel. Quantitative analysis of this data showed that the proliferation rate of unbranched lymphatic vessels approximated a linear mathematical function, about 1 cell/day, and that the proliferation rate of branched lymphatic vessels was more complex approximating a geometric function. We used low dose inductive conditions to label a trace frequency of LECs, a strategy that allowed us to visualize clonal expanding td tomato+ lineages within newly synthesized lymphatic vessels. These results suggest that multiple LECs contribute to sprout formation and that branch elongation results from LEC proliferation in a spatially restricted linear, rather than a circumferential, dimension.

Conclusions: The results are consistent with a progenitor model of corneal lymphangiogenesis in which many preexisting LECs support new lymphatic vessel growth in response to inflammatory disease conditions.

Keywords: 609 neovascularization • 480 cornea: basic science • 740 transgenics/knock-outs  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×