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Fig. 6. Spatial expression profile of enpp1, 4, 6 and 7 during development. Whole mount in situ hybridisation with DIG-labelled antisense RNA probes was performed on embryos from stages 5-41 for enpp1, 4, 6 and stages 32-45 for enpp7. All embryos were analysed from the same experimental set. The differences in colours result from clearing the embryos and photographing them against a white background to visualise staining. (A) Whole-mount in situ hybridisation analysis of enpp1 expression. Lateral view at stages 28 (i), 35/36 (ii) and 37/38 (iii). Detail of the fin at stage 41 (iv) and dorsal view of a stage 41 embryo (v). Anterior is right and dorsal is up, except in (v). da, dorsal aorta; dc, duct of cuvier; dlav, dorsal longitudinal anastomosing vessel; h, heart; pcv, posterior cardinal vein; ps, pronephric sinus; vbi, ventral blood island. (B) Whole-mount in situ hybridisation analysis of enpp4 expression. Animal view at stage 5 (i), anterior view at stage 22 (ii), lateral view at stages 32 (iii), 37/38 (iv) and 41 (v). For the lateral views, dorsal is up and anterior is right. Transverse sections at stage 37/38 through the proximal tubules (vi). Dorsal is up. The dotted line through the pronephric tubules (iii) correspond to planes of section in (vi). cg: cement gland; ct, collecting tubules; dt, distal tubules; hg, hatching gland; pt, proximal tubules; s, somites. (C) Whole-mount in situ hybridisation analysis of enpp6 expression. Lateral view at stage 5 (i), lateral view at stage 28 (ii) and 37/ 38 (iii). Dorsal is up and anterior is right. Transverse sections in the proximal tubules level at 37/38 (iv). The dotted line through the pronephric tubules correspond to planes of section in (iv). p, pronephros; pt, proximal tubules. (D) Whole-mount in situ hybridisation analysis of enpp7 expression. Ventral view of stage 45 embryos hybridized with anti-sense (i) and sense (ii) probe, lateral view of the gut region of a stage 45 embryo (iii), cryostat section (iv) at 28μm of a stage 45 embryo through the gut. no, notochord; m, muscles; sc, spinal cord; sia, first sections of the small intestine; sib, external coil of small intestine; sic, internal coil of small intestine (from Chalmers and Slack, 1998).
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Fig. 7. Comparison of the spatial expression profile of enpp2a [enpp2.S] and enpp2b [enpp2.L] genes during development. (A) Whole-mount in situ hybridisation with an enpp2a and enpp2b DIG labelled anti-sense RNA probe was performed on embryos from stages 5-41. Animal view at stage 5 (i, ii). Lateral view at stage 21 (iii, iv), stage 26 (v, vi), stage 32 (vii, viii) and stage 37/38 (ix, x). Dorsal is up and anterior is left. The dotted line through the fins (ix) corresponds to plane of section in Ci. (B) Whole-mount in situ hybridisation with an enpp2a and enpp2b DIG labelled antisense RNA probe on stage 32 embryos. Transverse section through the head (i, iv) and trunk (ii, v) levels. Lateral view of the trunk region (iii, vi). Arrowheads indicate the staining in the spinal cord. (C) Whole-mount in situ hybridisation with an enpp2a DIG labelled antisense RNA probe. Transverse section through the fin at stage 37/38 (i). Dorsal view of a stage 41 cleared embryo (ii). ba, branchial arches; df, dorsal fin; dt, distal tubules; e, eye; f, fin; fb, forebrain; fcr, fin crest; fco, fin core; h, heart; lh, lymphatic heart; lv, liver diverticulum; n, notochord; nc, neural crest; np, neural plate; ns, nasal placode; ov, otic vesicle; p, pharynx; pt, pronephric tubules; s, somites; sp, spinal cord; vf, ventral fin.
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enpp2 (ectonucleotide pyrophosphatase/phosphodiesterase 2) gene expression in Xenopus laevis embryos, NF stage 26, as assayed by in situ hybridization, lateral view, anterior left, dorsal up.
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enpp4 (ectonucleotide pyrophosphatase/phosphodiesterase 4) gene expression in Xenopus laevis embryos, NF stage 32, as assayed by in situ hybridization, lateral view, anterior right, dorsal up.
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enpp6 ( ectonucleotide pyrophosphatase/phosphodiesterase 6 ) gene expression in Xenopus laevis embryos, NF stage 37/38, as assayed by in situ hybridization, lateral view, anterior right, dorsal up.
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enpp7 ( ectonucleotide pyrophosphatase/phosphodiesterase 7 ) gene expression in Xenopus laevis embryos, NF stage 45, as assayed by in situ hybridization, ventral view.
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enpp1 (ectonucleotide pyrophosphatase/phosphodiesterase 1) gene expression in Xenopus laevis embryos, NF stage 41, as assayed by in situ hybridization,(iv) lateral view, anterior right, dorsal up; and dorsal view anterior right (v).
dlav: dorsal longitudinal anastomosing vessel ; da: dorsal aorta; pcv: posterio cardinal vein, ps: pronephric sinus.
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Figure 6 D. Spatial expression profile of enpp1, 4, 6 and 7 during development.Whole-mount in situ hybridisation analysis of enpp7 expression. Ventral view of stage 45 embryos hybridized with anti-sense (i) and sense (ii) probe, lateral view of the gut region of a stage 45 embryo (iii), cryostat section (iv) at 28μm of a stage 45 embryo through the gut. no, notochord; m, muscles; sc, spinal cord; sia, first sections of the small intestine; sib, external coil of small intestine; sic, internal coil of small intestine (from Chalmers and Slack, 1998).
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Ventral view of a X. laevis tadpole, NF stage 45, showing the external and internal coils of the small intestine.
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Nasal or olfactory placode (ns) and lymph hearts (left and right) (lh) are visible in this dorsal view of a NF stage 41 X. laevis tadpole, visualised via in situ hybridization for expression of enpp2a (ectonucleotide pyrophosphatase/phosphodiesterase 2).
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Suppl Table 1B. ACCESSION NUMBERS OF THE DIFFERENT CLONES USED IN THIS STUDY
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