Xenbase Image ID: 23281

Fig. 3. Wee1 depletion inhibits morphogenesis but not zygotic gene expression. (A) Animal cap explants prepared from uninjected embryos and embryos injected with MO-Control, MO-Wee1 or MO-Wee1+WT Wee1 RNA were left untreated or were treated with activin and cultured until stage 22-23. (B) RNA was isolated from animal cap explants prepared as in A and from stage 10.5 whole embryos (WE) that were either uninjected or had been injected with Control-MO or MO-Wee1 at the two-cell stage. Expression of brachyury, goosecoid and chordin was examined by RT-PCR analysis. cDNA levels were normalized to EF-1α, and a sample lacking reverse transcriptase (–RT) was also included. (C) Two-cell embryos were injected with MO-control, MO-Wee1 or MO-Wee1 + WT RNA. β-Gal RNA was injected into the B1 blastomeres at the 32-cell stage and β-gal activity visualized at stage 11.5-12. The B1 clone forms a narrow midline band extending between the blastopore (bottom) and animal hemisphere (top) in uninjected (n=12) and MO-Control injected embryos (n=23), while the B1 progeny form a broad band across the dorsal equator in MO-Wee1 embryos (MO-Wee1; n=36). this defect is significantly reversed by co-injection of WT Wee1 RNA (MO-Wee1+WT RNA; n=16). (D) The embryos shown in C were bisected through the area of β-gal staining. In uninjected embryos, the labeled cells extend from the animal hemisphere (top) to the dorsal blastopore lip (dbl). In Wee1-depleted embryos (MO-Wee1), no epibolic spread towards the vegetal pole (bottom) or involution occurs. However, some of the inner vegetal cells have moved upwards along the inner surface of the blastocoel roof (b.c.; arrow heads). (E) Expression of Xbrachyury (upper two panels, MO-Wee1, n=67) and chordin (MO-Wee1, n=54, lower two panels) was determined by in situ histochemistry (blue staining). Image published in: Murakami MS et al. (2004) Copyright © 2004. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue tbxt.S bra, brachyury, ntl, t, t-a, t-b, X-bra, Xbra, Xbrachyury X. laevis Throughout NF stage 10.5 marginal zone ventral marginal zone dorsal marginal zone involuting marginal zone mesoderm chrd.S chd, chordin, chrd.1, LOC108716939, X-chordin X. laevis Throughout NF stage 10.5 involuted dorsal mesoderm involuting marginal zone dorsal marginal zone Image source: Published Larger Image Printer Friendly View

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