Figure 3. Itgα8 is expressed periocular neural crest (pNC) during cornea development and plays a role in cell migration.

(A) Expression of Itgα8 is observed in pNC prior to their migration into the cornea at embryonic day (E)4 (arrow). (B) Itgα8 is subsequently expressed in the corneal endothelium at E5 (arrows) and (C) the migratory pNC in the corneal stroma at E6. (D) Schematic showing the isolation of periocular mesenchyme used for generating pNC explants for in vitro migration on Npnt-coated substrate in the presence or absence of α8β1 inhibitor. (E) Explant cultured on Npnt substrate showing robust cell migration after 12 hr. (F) Explant cultured on Npnt substrate in the presence of α8β1 inhibitor showing fewer cell migration after 12 hr. (G) Statistical analysis performed on N = 6 explants on Npnt substrate and N = 6 explants on Npnt substrate plus inhibitor revealed significant reduction in cell density of migratory cells in the presence of the inhibitor. *p<0.05. ec, ectoderm; pom, periocular mesenchyme; en, corneal endothelium; ep, corneal epithelium; st, stroma. Scale bars: 100 μm.

Figure 3—video 1. Migration of periocular neural crest (pNC) from mesenchyme explant on nephronectin (Npnt)-coated substrate.

Download video file ^{(12.1MB, mp4)}

Time-lapse movie was taken over 17 hr with images taken every 3 min and 27 s. The cells are shown in bright-field and fluorescent Hoechst nuclear staining (blue). Relates to Figure 3E.

Figure 3—video 2. Migration of periocular neural crest (pNC) from mesenchyme explant on nephronectin (Npnt)-coated substrate in the presence of α8β1 inhibitor.

Download video file ^{(8.9MB, mp4)}

Time-lapse movie was taken over 17 hr with images taken every 3 min and 27 s. The cells are shown in bright-field and fluorescent Hoechst nuclear staining (blue). Relates to Figure 3F.