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FIG. 10. Suppression of the anterior neural development in cocultures of ectoderm and early DMZ cells by blocking FGF signaling in ectoderm cells with DXFGFR-4a. (A) Schematic illustration of the experimental design. DXFGFR-4a mRNA was injected into four animal blastomeres at the eight-cell stage (150 pg/blastomere). The injected and uninjected embryos were incubated until stage 10, when ectoderm cells were isolated. A fixed number of these (160 cells/well) were cocultured in microculture wells with increasing number of DMZ cells (0 to 80 cells) prepared from intact early gastrula (stage 101/4). The transcription levels of four anterior neural markers, BF-1, Rx-1, XeNK-2, and En-2, and an epidermal marker, Keratin, in these cultures were analyzed as in Fig. 7. (B) Suppression of the neural-inducing and epidermal-inhibiting action of early gastrula DMZ (organizer) cells by overexpression of DXFGFR-4a in ectoderm cells. Autoradiographs are shown of RT-PCR products of the marker transcripts coamplified with EF1a transcript in cocultures including uninjected (left panels) and injected (right panels) ectoderm cells. The data from cultures of ectoderm cells (160 cells/well) or DMZ cells (80 cells/well) alone are also included. (C) Quantitative assessment of the effects of overexpressing DXFGFR-4a. Each RT-PCR product shown in (B) was quantified as in Fig. 5B. Values are normalized as in Fig. 7B and plotted against cocultured DMZ cell numbers (0 to 80 cells). Image published in: Hongo I et al. (1999) Copyright © 1999. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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