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FIG. 5. Detailed expression pattern analysis of P7E4, P8F7, and P17F11 by whole-mount in situ hybridization. Whole or hemisectioned embryos were hybridized with P7E4 (A), P8F7 (G), P17F11 (L), or crescent (Q, R) probe. Other halves of hemisectioned embryos were used for dkk-1, cerberus (cer), chordin (chd), or XHex probe as indicated. Stained embryos were treated with BB/BA to become transparent for observation (B, F, K, P, right panel). Sections were made from stained whole embryos (C, H), and are shown in bright field and with DAPI staining. (A) P7E4 (homologous to human hypothetical proteins, BDPL and KIAA0952). (A) Sagittal hemisections of early gastrulae (stage 10.5) stained for P7E4 (upper panel) and dkk-1 (lower panel). P7E4 expression was detected around the Brachet cleft (arrowhead) and weakly in the mesoderm region (arrow). Dorsal to the right. (B) Dorsal view of early neurula. Anterior to the left. Cross sections were made at the indicated positions (C) which correspond to the following panels (C). (C) Sections of the same embryo as shown in (B). P7E4 was expressed in the archenteron roof as well as the bilateral domains including the neuroectoderm and mesoderm region (arrowheads) (C). The bilateral expression domains were notable in the mesodermal region at position D. In the posterior region, expression was seen in the notochord (E). (F) Dorsal view of sagittally hemisectioned neurula embryos stained for P7E4 and chordin. Anterior is up. (G) P8F7 (unknown). (G) Sagittal hemisections of early gastrula stained for P8F7 (top) and dkk-1 (middle), and vegetal view of the hemisectioned embryo (bottom). Right panels, higher magnification of the left panels. Dorsal is to the right (top, middle) or up (bottom). Arrow, the surface layer of the dorsal blastopore lip, expressing P8F7 but not dkk-1; arrowhead, punctuated expression of P8F7 in the epithelial layer of the dorsal ectoderm. (H) Vegetal view of early gastrula (upper panel, dorsal is up) and a sagittal section (lower panels, dorsal to the right). Arrowhead, dorsal blastopore lip; arrow, anterior archenteron. (I) Sagittal hemisections of midgastrula stained for P8F7 and chordin. Dorsal to the right. Arrowhead, expression of P8F7 in the anterior archenteron roof. (J) Sagittal hemisections of early neurula stained for P8F7 and XHex. Anterior to the left. Arrow, expression in the anterior endoderm; arrowhead, expression in the anterior archenteron (P8F7) or the prechordal plate (XHex). (K) Dorsal view of sagittally hemisectioned neurula stained for P8F7 and chordin. Anterior is up. (L) P17F11 (related to human MGC13045 and Arabidopsis CAB90947). (L) Sagittal hemisections of late blastula stained for P17F11 and chordin. (M) Sagittal hemisections of early gastrula stained for P17F11 (top) and dkk-1 (middle), and vegetal view of the hemisectioned embryo (bottom). Dorsal is to the right (top, middle), or up (bottom). Arrowhead, the surface layer of dorsal blastopore lip, expressing P17F11 but not dkk-1. (N, O) Sagittal hemisections of late gastrulae stained for P17F11, chordin, or XHex. Dorsoanterior to the right. (P) Sagittal hemisections of early neurula stained for P17F11 (upper) and chordin (lower), and dorsal view of the hemisectioned embryo (right). Partial breakage of the archenteron roof shows interior staining in the anterior mesoderm as indicated by af. Anterior is to the left (left panels), or up (right panel). (Q, R) crescent. (Q) Sagittal hemisections of early gastrula stained for crescent (upper) or dkk-1 (lower), and vegetal view of the hemisectioned embryo (right). Dorsal is to the right (left panels), or up (right panel). (R) Sagittal hemisections of early gastrula stained for crescent and cerberus. Dorsal is to the right. Arrowheads, the surface layer of dorsal blastopore lip, expressing crescent but not dkk-1 or cerberus. Numerical figures in the panels indicate the developmental stages according to Nieuwkoop and Faber (1967). Pictures of hemisections stained for dkk-1, cerberus, chordin, and XHex were flipped for comparison. af, artifact; en, endoderm; nc, notochord; ne, neuroectoderm; sm, somitic mesoderm. |
