Fig. 1: Ocular Angpt1-expressing tissues are derived from the neural crest.
A Angpt1-GFP staining showing expression in PDGFRB-positive tissues of the uveal tract. As expected, no Angpt1 expression was detected in PODXL-positive endothelial cells. Panel shown is a representative section from a group of four littermate animals. B Visualized using the Rosa26mT/mG reporter mouse line, cre-mediated recombination in the neural crest lineage using Wnt1-Cre recapitulated Angpt1 expression in the trabecular meshwork and uveal tissues. In the retina, recombination was limited to pericytes (white arrows). C Neural crest specific knockout (Angpt1ΔNC) mice are born normally in the predicted Mendalian ratio. p = 0.56 as determined by two-tailed Fisher’s exact test. Pups weaned: Control, n = 78, Angpt1ΔNC, n = 66. D As measured in 12-week-old mice using quantitative rtPCR, Angpt1 mRNA expression was dramatically reduced in the limbus/iridocorneal angle region (LI) of Angpt1ΔNC mice (p = 0.0001). A moderate reduction in expression was observed in the retina (p = 0.0203), consistent with retinal Angpt1 expression by both NC and non-NC-derived cells. In contrast, Angpt1 expression in solid organs such as the lung was not altered (p = 0.368). Each point represents expression measured in an individual animal (two pooled eyes). Control, n = 5, Angpt1ΔNC, n = 7 (limbus) and control, n = 8, Angpt1ΔNC n = 11 (retina and lung). Scale bars represent 50 μm (A,B). *p < 0.05, ***p < 0.001 as determined by two-tailed Student’s t-test. DfLI = 10, Dfretina, lung = 17.
