The cellular bases of choroid fissure formation and closure

[摘要] Defects in choroid fissure (CF) formation and closure lead to coloboma, a major cause of childhood blindness. Despite genetic advances, the cellular defects underlying coloboma remain poorly elucidated due to our limited understanding of normal CF morphogenesis. We address this deficit by conducting high-resolution spatio-temporal analyses of CF formation and closure in the chick, mouse and fish. We show that a small ventral midline invagination initiates CF formation in the medial-proximal optic cup, subsequently extending it dorsally toward the lens, and proximally into the optic stalk. Unlike previously supposed, the optic disc does not form solely as a result of this invagination. Morphogenetic events that alter the shape of the proximal optic cup also direct clusters of outer layer and optic stalk cells to form dorsal optic disc. A cross-species comparison suggests that CF closure can be accomplished by breaking down basement membranes (BM) along the CF margins, and by establishing BM continuity along the dorsal and ventral surfaces of the CF. CF closure is subsequently accomplished via two distinct mechanisms: tissue fusion or the intercalation of various tissues into the inter-CF space. We identify several novel cell behaviors that underlie CF fusion, many of which involve remodeling of the retinal epithelium. In addition to BM disruption, these include NCAD downregulation along the SOX2+ retinal CF margin, and the protrusion or movement of partially polarized retinal cells into the inter-CF space to mediate fusion. Proximally, the inter-CF space does not fuse or narrow and is instead loosely packed with migrating SOX2+/PAX2+/Vimentin+ astrocytes until it is closed by the outgoing optic nerve. Taken together, our results highlight distinct proximal-distal differences in CF morphogenesis and closure and establish detailed cellular models that can be utilized for understanding the genetic bases of coloboma.

[学科分类] 生物科学（综合）

[关键词] A;anterior;BM;basement membrane;CF;Choroid Fissure;D;dorsal;Di;distal;E;embryonic;Fib;fibronectin;HA;hyaloid artery;HH;Hamburger and Hamilton embryonic stages;hpf;hours post fertilization;ILM;inner limiting membrane;L;lens;LAM;laminin;M;medial;OC;optic cup;OD;optic disc;OL;outer layer of the optic cup;ON;optic nerve;OS;optic stalk;OV;optic vesicle;P;posterior;Pec;pecten;Phal;phalloidin;pHH3;phospho-histone H3;POM;periocular mesenchyme;Pr;Proximal;Pyr;Pyronine Y;R;retina;RPE;retinal pigment epithelium;Vim;Vimentin;Coloboma;Optic cup morphogenesis;Optic disc;Optic nerve;Optic fissure;Tissue polarity and epithelial reorganization;Tissue fusion;Pecten