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Fig. 4. Whole mount in situ hybridisation of Pod 1 in X. laevis. Wholemount in situ hybridisation with a Pod 1 DIG-labelled antisense (A,C,D,F,G and I) and sense (B,E,H and J) RNA probe was performed on embryos at the stage indicated. Embryos in panels D, G, I and J have been cleared in Murray's solution to facilitate the observation of internal staining. No staining above background was present in embryos at stage 22 (A,B) as staining observed in the eye placode, ep, and somites, s, was present in the antisense and sense in situ hybridisations. (C,D) Glomus, g and anterior branchial arch, b, staining is visible in uncleared and cleared stage 28-29 embryos. (E) Stage 29 sense RNA probed embryos do not show staining in the branchial arch or the glomus. (F) Uncleared antisense stage 33 embryos show discrete staining in the branchial arches, b and the glomus, g, which is also observed in the cleared embryos (G). (H) Sense stage 33 embryos also have some weak staining in the branchial arches. The staining observed in the eye is observed in both antisense and sense stage 33 embryos and is considered to be background (F,H). The late tailbud embryos are stained in the epicardium, h, of the heart and in all three components of the pronephros, the glomus, g, tubules, t and duct, d (I). Non-specific staining is seen in the anterior region of late stage embryos in both sense and antisense hybridised embryos (J). |