September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Quantitative assessment of vessel regression and pericyte detachment in a mouse model of retinal angiogenesis
Author Affiliations & Notes
  • Sabine Uhles
    Pharma Research and Early Development, Ophthalmology Discovery and Biomarkers, Roche Innovation Center Basel, F. Hoffmann - La Roche Ltd., Basel, Switzerland
  • Franco Revelant
    Pharma Research and Early Development, Ophthalmology Discovery and Biomarkers, Roche Innovation Center Basel, F. Hoffmann - La Roche Ltd., Basel, Switzerland
  • Sabine Grüner
    Pharma Research and Early Development, Ophthalmology Discovery and Biomarkers, Roche Innovation Center Basel, F. Hoffmann - La Roche Ltd., Basel, Switzerland
  • Guido Hartmann
    Pharma Research and Early Development, Ophthalmology Discovery and Biomarkers, Roche Innovation Center Basel, F. Hoffmann - La Roche Ltd., Basel, Switzerland
  • Fethallah Benmansour
    Pharma Research and Early Development, pRED Informatics, Roche Innovation Center Basel, F. Hoffmann - La Roche Ltd., Basel, Switzerland
  • Footnotes
    Commercial Relationships   Sabine Uhles, F. Hoffmann - La Roche Ltd. (E); Franco Revelant, F. Hoffmann - La Roche Ltd. (E); Sabine Grüner, F. Hoffmann - La Roche Ltd. (E); Guido Hartmann, F. Hoffmann - La Roche Ltd. (E); Fethallah Benmansour, F. Hoffmann - La Roche Ltd. (E)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4534. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Sabine Uhles, Franco Revelant, Sabine Grüner, Guido Hartmann, Fethallah Benmansour; Quantitative assessment of vessel regression and pericyte detachment in a mouse model of retinal angiogenesis. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4534.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The neonatal mouse retina has become a prominent model for studying angiogenesis. However most quantitative assessments of anti-angiogenic therapeutics inhibiting vascular growth, sprouting and maturation are performed on randomly selected fields of microscopic view that introduce a high risk of biased measurement. We aimed to develop a semi-automated image analysis solution for annotation and segmentation of flat-mounted retinal scans, enabling computation of large specimen numbers and differentiation between applied anti-angiogenic modalities.

Methods : Neonatal mice were treated i.p. at day of birth, P2 and P4 with anti-angiogenic compounds influencing vascularization and pericyte (PC) attachment. Pups were sacrificed on P5. Retinal flatmounts, stained for endothelial cells (EC) and PCs, were imaged with an automated scanner (Olympus) and subjected to quantitative image analysis employing an algorithm for vessel growth and pericyte attachment (Definiens XD). Quantification of the superficial vascular plexus and EC coverage by PCs was analyzed by segmentation and analysis of the digitized data per gray-scale analysis followed by channel co-localization. Parameters to differentiate anti-angiogenic compounds were radial expansion of vessel growth, % vascular plexus area, branching, endothelial tip cells, vascular front tortuosity, and % PC-covered vessels.

Results : Analyses were performed on wing-based computation and data visualized using Tibco Spotfire. Results significantly differentiated the effects of single receptor antagonists as well as their monomeric combinations and revealed comparable and statistically relevant regression on vascular growth and maturation. Quantification of PCs attached to annotated retinal arterioles vs venules vs capillaries revealed clear differences between the vascular beds. However a statistical compound effect was only detectable on the venules.

Conclusions : We report generation of an image analysis solution that allows semi-automatic quantification and differentiation of anti-angiogenic therapeutics in a mouse model of angiogenesis. This unbiased approach delivers significant and reproducible results over a series of studies and provides a promising alternative to manual procedures.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

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.

×