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Anti-runx2 morpholino injections do not affect cranial neural crest cell migration. A and C are lateral views of live stage-33/34 hosts that have received neural crest transplants. B and D are the stage-44 hatchlings of the same individuals as A and C, which have been stained with the anti-collagen II antibody (red in B, white in D). A: Live image of a wild-type host that has received neural crest from a morpholino-injected donor. Portions of the presumptive hyoid and branchial neural crest streams have been replaced with morpholino-injected cells (green), which are migrat- ing normally. B: Same individual as A at stage 44, stained with anti-collagen II to reveal cartilage formation (red). The ceratohyal cartilage is truncated on the transplanted side, with a population of donor cells derived from the hyoid stream concentrated on the distal end (arrow). C,D: Neural crest transplanted from a wild-type donor into a morpholino-injected host can rescue cartilage loss. C: Live image of a morpholino-injected host that has received wild-type neural crest. The rhodamine labeled wild-type crest (red) migrates normally in the morpholino-injected host (green). D: Same individual as C at stage 44, stained with anti-collagen II (white). The ceratohyal is partially rescued on the morpholino-injected side (arrow). The rhodamine-label of the wild-type crest did not persist through antibody staining. E,F: Sox9 in situ hybridization in a single stage-32 embryo unilaterally injected with both runx2-I and runx2-II isoform-specific morpholinos. E: Wild-type side of the embryo, showing normal neural crest morphology revealed by the distribution of the sox9 probe. F: Anti-runx2-morpholino-injected side showing the same sox9 staining pattern as the wild-type side. The fluorescein label did not persist through the whole mount in situ hybridization. Abbrevi- ations as in Figures 2 and 4. |
