Cousin H .

R03 DE025692 NIDCR NIH HHS

Fig. 1. Origin and migration of the cranial neural crest cells in Xenopus laevis. (A) Schematics representing transverse sections though the prospective otic placode (Ot) of Xenopus embryos at the various stages of development ( Nieuwkoop and Faber, 1967). Dorsal is up. The placodal ectoderm and neural crest prospective tissue emerge from the sensory layer of the ectoderm and are anatomically visible as soon as stage 14 (early neurulation). At stage 19 (late neurula stage) the cranial neural crest starts migrating ventrally (Sadaghiani and Thiebaud, 1987). (B) Schematic showing the relative positioning of CNC and the cranial placodes ( Sadaghiani and Thiebaud, 1987 ; Schlosser, 2006). During the early neurula stage (stage 15) both placodal (blue) and CNC (pink) primordium appose one another. Starting with the early tailbud stage (stage 21), these territories are segmented into various subgroups that reflect their future differentiation. The placodal ectoderm secretes Sdf1, which attracts CNC. Once the CNC reaches a placode, CIL takes place and the placode migrates away (i.e. ventrally). Once the placodes and CNC are separated, the Sdf1-based chemoattraction reasserts itself. This “chase and run” contributes to the proper migration of the each of the CNC segments while the placodes lying in the path of the CNC (e.g. Epibranchial placodes) reach their proper dorsoventral location and are shaped into narrow strips of tissues. Ot: Placode; NT, neural tube; n: notochord; psm: presomitic mesoderm, so: somites; LPM: lateral plate mesoderm, CG: cement gland; Op: optic vesicle.

Fig. 2. Migration of CNC cells in vitro and in vivo. GFP expressing CNC are explanted at stage 15 and either grown on a substrate of fibronectin in a saline media (A) or grafted into a wild type embryo (B) (anterior left, dorsal top). The migration of the CNC is followed by time-lapse photography (3 minute intervals, 10 h of total migration) and still pictures from these movies were extracted at regular time points throughout the movies. The migration of CNC in vitro (A) recapitulates the migration of the CNC observed in vivo (B) including the formation of segments (mandibular, hyoid, branchial), and the collective migration which is observed in the first 3 frames and the single cell migration which is observed in the last frame. Note that the explant in A was cut purposely large to demonstrate the formation of segments and therefore contains some neural tissue contaminant (NT). m: mandibular; h: hyoid segment b: branchial segments (b, b1, b2); CG: cement gland.

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