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Figure 1. eg5 expression pattern in developing Xenopus embryos. Whole-mount in situ hybridization analysis of spatio-temporal eg5 expression. A–D: Dorsal view, anterior on the left. E: Dorso-lateral view. H, J: Lateral view, anterior on the left, dorsal at the top. F, G, I: Transversal sections. Dashed lines in C, E, and H indicate the sites of transverse section shown in F, G, and I, respectively. A: eg5 transcripts are first detected since the late gastrula stage (stage 12.5) in the dorsal region of embryos at the neural plate and neural plate borders (arrowheads). B–D: During neurulation (stages 14–18), eg5 is expressed in the neural plate borders and in prospective neural crest (arrowheads). No expression is detected in the neural plate midline (arrow). The transversal section of a stage-16 midneurula embryo (F) shows eg5 expression in neural plate (black bracket) and in the neural crest region (red brackets). The transversal section of stage-21 embryo (G) shows eg5 transcripts in the dorsal region of dermatomes (arrowheads). No specific labeling is observed in the sense probe control (C, inset). E: Stage-21 embryos show eg5 expression in the head region, Rohon-Beard sensory neurons (arrows), and the epidermis in a spotted pattern. H: Stage-24 embryos show eg5 expression at the somites, optic vesicle, profundal ganglia primordium (encircled), and epidermis external layer (I). J: The tailbud-stage embryos show the expression at the branchial arches, somites, brain, and epidermis. n, notochord; nt, neural tube; s, somites; b, brain; ba, branchial arches; op, optic vesicle. K–M: Double in situ hybridizations for eg5 (purple) and foxd3 (turquoise), sox2 (turquoise), and xk81a (turquoise). Dorsal view, anterior on the left. K'–M': Transverse sections of St. 15 neurula embryos displayed in K–M showing that eg5 expression overlaps with foxd3 in the neural crest territory (K', black brackets: foxd3 neural crest expression), and is complementary with sox2 and xk81a expressions (L', M'). L': Red brackets, neural crest region; black bracket, neural plate. M': Black brackets, neural crest region. N–P: RT-PCR analysis of eg5 expression in developing embryos. O: Quantification of the gel shown in N, the results are expressed as Relative intensity (sample/ef1α x10). P: RT-PCR analysis of eg5, foxd3, sox2, myod and xk81a expression in neural crest, intermediate mesoderm, neural plate and non-neural ectoderm explants. Explants were dissected out from stage-14 embryos (see Experimental Procedures section). ef1α, loading control. np, neural plate; n, notochord; nc, neural crest; m, intermediate mesoderm; nne, nonneural ectoderm; ac, animal cap (Stage 9).Download figure to PowerPoint |
