The Use of Multidimensional Microscopy in the Characterization of Retinal Vascular Development in a Murine Model

Abstract

The murine retinal vasculature develops in its entirety following birth through the process of angiogenesis. This developing system provides an extremely accessible model for investigation of angiogenic mechanisms in general and as they pertain to vascular diseases of the eye (e.g. diabetic retinopathy and macular degeneration). The model is particularly relevant clinically because the developmental events of retinal vascularization are nearly identical in mice and humans, although primate vessels develop in utero during the final trimester of pregnancy and are thus less accessible for experimental study. The model is also excellent for the testing of anti-angiogenic agents as potential treatments for vascular diseases. in this report we outline the use of confocal optical microscopy and multiphoton microscopy in the 3-dimensional characterization of retinal vascular development in the mouse.

We employed confocal microscopy to characterize the events of vascular development from postnatal days 0-56 (P0-P56). During this time the retinas are transformed from simple, thin bilaminar structures to more complex, functional, adult sensory organs. Retinal morphogenesis continues until P42. Retinas were dissected from eyes and vessels stained with collagen type-IV antibody (1) or Bandeiraea simplicifolia (Griffonia) lectin-dye conjugate (2) to outline the blood vessels. All patent vessels were stained by both techniques.