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Fig. 6. EZH2 regulates neural crest migration. (A) Wild-type embryos were cultured in medium with DMSO or 50 µM EZH2 inhibitor GSK126 from stages 13-14 onward. The embryos were analyzed by ISH for Sox10 and Twist expression at tailbud stages 25-26. In DMSO-treated embryos, Sox10- and Twist-expressing cells migrated in distinct streams in the head. Treatment with GSK126 resulted in reduced migration of these cells. In addition, Sox10 and Twist expression in dorsal midline of the trunk (arrows) was eliminated by GSK126 treatment. (B) Neural crest tissues from fluorescein dextran-labeled donor embryos were transplanted into unlabeled control embryos at neurula stages 15-16. The embryos were cultured in medium with DMSO or 50 µM GSK126 until tailbud stages 26-28. Migration of the neural crest was examined by fluorescence microscopy. Whereas neural crest from DMSO-treated embryos migrated efficiently in the head, GSK126-treated embryos displayed defects in neural crest migration. (C) Neural crest explants were dissected from stage 15-17 embryos and cultured on fibronectin-coated dishes for about 16 h in the presence of DMSO or 8-10 µM GSK126. Migration of the neural crest was imaged between 2 and 16 h of culture. Whereas neural crest cells dissociated from the core explants and migrated efficiently in DMSO-containing medium, treatment with GSK126 reduced the migration of the neural crest on fibronectin. (D) Tracking of cell migration in time-lapse movies revealed that the velocity of neural crest cell migration was significantly reduced by treatment with GSK126. Four movies each for DMSO- and GSK126-treated samples were analyzed, with 60-170 cells tracked in each movie. |
