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Figure 3. In situ hybridization of Mig30. Positions of sections are indicated by dotted lines, and letters mark corresponding panels. A-C: Sagittal sections; dorsal to the left. A: Stage (st.) 8.5; asterisk marks the blastocoele chordin- and noggin-expressing center. B,C: Black arrowheads mark the dorsal lip. B: St. 10.25. C: St. 11.5. D: St. 14; anterior view. E-G: St. 17. E: Anterior view. F: Dorsal view, anterior to the left. G,H: Transverse sections. I,J: St. 20. I: Anterior view. J: Dorsal view, anterior to the left. K,L: St. 31. K: Lateral view. L: Transverse section. M,M prime : St. 39. M: Lateral view. M prime : Transverse section. bt, branchial tissue; e, eye; he, heart; le, lens; lp, prospective lens placode; no, notochord; nt, neural tube; otp, prospective otic placode; otv, otic vesicle; re, retina; so, somite.
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Figure 4. In situ hybridization of xIGFBP-rP10. Positions of sections are indicated by white dotted lines and letters mark corresponding panels. A,B: Sagittal sections. A: Stage (st.) 4. B: St. 10.5; black arrowheads mark dorsal lip; insert: vegetal view. C,D: St. 15. C: Anterior view. D: Dorsal view, anterior to the left; insert: transverse section. E,F: St. 21. E: Anterior view. F: Dorsal view, anterior to the left. G-I: St. 24. G: Lateral view; insert: vegetal view of head. H: Transverse section. I: Crude horizontal section at the level of the notochord. J-M: St. 30. J: Lateral view; insert: vegetal view of head. K-M: Transverse sections. N,O: St. 35; lateral view. O: Magnification of head region. P,Q: St. 40; lateral view. O: Magnification of head region; reversed saturation image of P, to visualize pattern of strongly stained areas. cg, cement gland; cnh, chordoneural hinge; df, dorsal fin; fp, floor plate; hm, head mesenchyme; no, notochord; otv, otic vesicle; so, somite; vf, ventral fin; VII and IX/X, cranial ganglia.
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Figure 5. Whole-mount in situ hybridization of xIGFBP-like. Positions of sections are indicated by white dotted lines, and letters mark corresponding panels. A,B: Stage (st.) 15. A: Anterior view. B: Dorsal view, anterior to the left. C: St. 18; anterior view. D-G: St. 21. D: Anterior view. E: Dorsal view, anterior to the left. F: Reversed saturation image of D, to visualize pattern of strongly stained areas. G: Transverse section. H-J: St. 25. H: Lateral view. I: Dorsal view, anterior to the left. J: Transverse section. K-M: St. 32. K: Lateral view. L,M: Transverse sections. N-Q: St. 39. N: Lateral view. O: Magnification of head region, dorsal view, anterior to the left. P: Transverse section. Q: Transverse section. aen, archencephalic region; anp, anterior neural plate; er, prospective eye region; fb, forebrain; hb, hindbrain; le, lens; mb, midbrain; nr, neural retina; no, notochord; otv, otic vesicle; ov, optic vesicle; pnr, prospective neural retina; psm, presomitic mesoderm; sc, spinal cord; so, somite.
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mig30 (mig30) gene expression in Xenopus laevis embryo, as assayed by in situ hybridization, NF stage 39, lateral view, anterior left, dorsal up.
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igfbpl1 (insulin-like growth factor binding protein-like 1) gene expression in a Xenopus laevis embryo, as assayed by in situ ybridization, NF stage 41, lateral view, anterior left, dorsal up.
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Figure 1. A: Schematic diagram of the protein domain architecture. Black, signal peptide (SP); yellow, insulin growth factor-binding homologues (IB, SMART accession no. 00121); blue, kazal-type serine protease inhibitors (KAZAL, SMART accession no. 00280); green, overlap of IB and KAZAL; orange, immunoglobulin C-2 type (IGc2, SMART accession no. 00408). B: Sequence alignment of hIGFBP-rP1 (GenBank accession no. NP_001544), xIGFBP-like (GenBank accession no. DQ515890), and xIGFBP-rP10 (GenBank accession no. AAH72910, Xenopus laevis MGC80370 protein) together with their human homologues (hIGFBP-like, GenBank accession no. NP_001007564; hIGFBP-rP10, GenBank accession no. NP_112191) and Mig30 (GenBank accession no. BAB84702). Note: The xIGFBP-like is not a full-length sequence. Conserved amino acids are shaded in gray. Dashes indicate gaps introduced to maximize the alignments. Protein domains are highlighted with same colors as in A. Note: The white box in the overlap of IB and KAZAL of hIGFBP-rP10 and white box in IB of xIGFBP-rP10 mark single amino acid substitutions (see the Results section). C: Cladogram of human and Xenopus laevis IGFBPs and IGFBP-rPs (see Table 1). The proteins characterized in this study (marked with a dot) together with their human homologues and hIGFBP-rP1 form a group (shaded in gray). For the putative Xenopus homologues of hIGFBP-1 and -2, GenBank accession numbers are AAH60008 and AAH84133, respectively. The tree was generated using ClustalW.
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Fig. 2. Expression of Mig30, xIGFBP-like, and xIGFBP-rP10 during early Xenopus laevis embryogenesis determined by reverse transcription-
polymerase chain reaction. Developmental stages after Nieuwkoop and Faber (N.F.; Nieuwkoop and Faber, 1967) are indicated. Ornithine decarbox-
ylase (ODC) served as the loading control.
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mig30 (mig30) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 8.5, saggital section, anterior up.
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mig30 (mig30) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 10.5, saggital section, anterior up.
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mig30 ( mig30) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 17, anterior view, dorsal up (E) and dorsal view, anterior right (G).
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mig30 (mig30) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up (K), and a transverse section through trunk section, dorsal up (L).
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kazald1 (Kazal-type serine peptidase inhibitor domain 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 3 (4-cell), lateral view, anterior left, dorsal up.
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kazald1 (Kazal-type serine peptidase inhibitor domain 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 15, lateral view, anterior left, dorsal up.
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kazald1 (Kazal-type serine peptidase inhibitor domain 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anterior left, dorsal up.
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kazald1 (Kazal-type serine peptidase inhibitor domain 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 40, lateral view, anterior left, dorsal up.
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igfbpl1 ( insulin-like growth factor binding protein-like 1) gene expression in a Xenopus laevis embryo, as assayed by in situ ybridization, NF stage 15 lateral view, anterior left, dorsal up.
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igfbpl1 ( insulin-like growth factor binding protein-like 1) gene expression in a Xenopus laevis embryo, as assayed by in situ ybridization, NF stage 21, anterior view, dorsal up.
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igfbpl1 ( insulin-like growth factor binding protein-like 1) gene expression in a Xenopus laevis embryo, as assayed by in situ ybridization, NF stage 21, lateral view, anterior left, dorsal up (H), and dorsal view (I).
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igfbpl1 ( insulin-like growth factor binding protein-like 1) gene expression in a Xenopus laevis embryo, as assayed by in situ ybridization, NF stage 21, lateral view, anterior left, dorsal up (H), and dorsal view (I).
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