Presence of RPE-Produced VEGF in a Timely Manner Is Critical to Choroidal Vascular Development
Choroidal circulation is responsible for approximately 80% of blood supplies to the eye and its health is essential to visual function. To gain better understanding of choroidal vascular system, we (1) generated mice with inducible disruption of vascular endothelial growth factor (VEGF) in the retinal pigment epithelium (RPE) and (2) developed a semiquantitative assay to evaluate the density of choroidal vessels in these mice. In this chapter, we report an improvement to our assay by staining choroidal vessels through perfusion of fluorescently labeled concanavalin A. We will also review our investigation on the temporal role of the RPE-produced VEGF in choroidal vascular development in mice. Our study establishes a timeline that the presence of the RPE-produced VEGF from embryonic day 10 to 15 is critical to choroidal vascular development. Therefore, mice with induced disruption of the RPE-produced VEGF after organogenesis do not have apparent developmental defects and may have utilities in investigating postdevelopmental function of VEGF, which is critical to the understanding of the relationship between the RPE and choriocapillaris in the pathogenesis of age-related macular degeneration.
KeywordsDevelopment Choroidal vasculature VEGF RPE Inducible Knockout
This study was supported partially by American Health Assistance Foundation grant M2008-059, Foundation Fighting Blindness grant BR-CMM-0808-0453-UOK, Beckman Initiative for Macular Research Grant 1003, NIH grants R01EY20900, P20RR17703, P20RR024215, and P30EY12190. American Diabetes Association grant 1-10-BS-94, Oklahoma Center for Advancement of Science and Technology Contract HR09-058, and Unrestricted Research Awards from Hope for Vision and Research to Prevent Blindness.