| |
Figure 4. Xenopus Rho-associated kinase alpha (xROK alpha ) regulates convergent extension (CE) movements without affecting gene expression. A: Temporal expression pattern of xROK alpha . Developmental stages are shown above each lane. Ornithine decarboxylase (ODC) was used as a loading control. B-D: Spatial expression pattern of xROK alpha . B: Expression pattern of xROK alpha at a stage 10.25 gastrula (dorsovegetal view). An arrow indicates the dorsal lip of blastopore. C: Sagittal section of a stage 10.25 gastrula. xROK alpha transcripts were mainly expressed in the dorsal mesoderm and ectoderm above the dorsal lip. An arrow indicates the dorsal lip of blastopore. D: At the stage 35 tadpole, xROK alpha was expressed in head and tail ectoderm and mesoderm. E: Dorsal two blastomeres of four-cell stage embryos were injected with xROK alpha mRNA (500 pg) and allowed to develop to stage 33 (lateral view). Two blastomeres of four-cell stage embryos were injected dorsally with 750 pg of xROK alpha . Keller explants were dissected at stage 10.5 and cultured until siblings were reached at stage 19. WT, wild-type. F: Quantitation of relative rescue in whole embryos for E. G: Quantitation of relative elongation of Keller explants compared with controls for E. In the Student's t-test, all differences are statistically significant (P < 0.05). These examinations were performed more than three times. Others indicate truncated axis and mild kinked axis. H,I: Analysis of reverse transcriptase-polymerase chain reaction demonstrated that xROK alpha does not affect dorsal cell fate specification in Keller explants cultured to stage 12 (H) and dorsoventral patterning of the mesoderm in Keller explants cultured to stage 22 (I). W.E indicates whole embryo; -RT indicates no reverse transcriptase control. ODC is the loading control. |
