Figure 7

Spatiotemporal expression patterns of conserved corneal epithelium and endothelium associated genes. (A) WISH analysis of corneal epithelium associated genes zgc:92,380, sparc, pfn1, epcam and col1a1b expression at 24, 72 and 120 hpf displayed in dorsal and lateral views. (B) 10 µm cryosections of 120 hpf embryo eyes after WISH. (C) Two color FWISH depicting co-expression of krt4(red)/zgc:92,380(green), sparc(green)/krt4(red), pfn1(green)/krt4(red), epcam(green)/krt4(red) and col1a1b(green)/krt4(red) at 120 hpf. DAPI (blue) was used to stain the nuclei. Volume projections in lateral view and single section dorsal views from 3D confocal stacks are displayed. White arrows indicate example regions of co-expression. Scale bar = 100 µm. Individual fluorescence channels are provided in Fig. S10. (D) Average mean expression measurements from CEp1 and CEp2 clusters for marker genes krt4, krt5, krt91 and icn2 as well as col1a1b, epcam, pfn1, sparc and zgc92380 at each timepoint examined, 48–144 hpf. (E) WISH analysis of corneal endothelium associated genesmatn4 and cthrc1a expression at 24, 72 and 120 hpf displayed in dorsal and lateral views. (F) 10 µm cryosections of 120 hpf embryo eyes after WISH. (G) Average mean expression measurements from CEn clusters for pmp22a, ctnnb2, matn4 and cthrc1a at each timepoint examined, 48–144 hpf. ALP Annular ligament progenitor (yellow arrow), CEp Corneal epithelium (blue arrow), CEn Corneal endothelium (green arrow), CSt Corneal stroma (green arrow).