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Figure 8. Zfp36 mRNA overexpression alters the formation of pronephros and affects pronephric marker genes expression. (A) 250 pg of Xenopus zfp36 mRNA were injected into one ventral blastomere of 8-cell stage embryos and developing embryos were fixed at stage 39 before immunhistochemistry analysis with the pronephros specific markers 3G8 and 4A6. Arrows in b and d mark the pronephros (pn) alteration on the injected side. (B) 250 pg of Xenopus zfp36 (a) or zfp36l1 (e) mRNA was injected into one ventral blastomere of 8-cell stage embryos and developing embryos were fixed at stage 22 (a, b and e) or stage 26 (c, d) before in situ hybridization analysis for pax8 or lim1 expression. Arrows in b, d, f and h mark the pronephros (pn) alteration on the injected side. (C) Two-cell stage embryos were injected or not (NI) with 250 pg of the different Xenopus zfp36 mRNAs. Animal caps were explanted at stage 8.5 and treated with activin plus retinoic acid (RA) before analysis by RT-PCR for smp30 expression when control embryos reached stage 35. Stage 35 embryo (Emb) or untreated animal caps (-) were assayed by RT-PCR in parallel. Odc was used as control of loading and a reaction was performed in the absence of reverse transcriptase to check for genomic DNA contamination (-RT). |
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Figure 10. Zfp36 depletion does not affect somitogenesis nor early pronephros specification. (A) 20 ng of zfp36 morpholinos (MO) were injected into one dorsal blastomere of 4-cell stage embryos with 250 pg of lacZ mRNA. Embryos were fixed at stage 15 for the detection of myod by in situ hybridization (a) or at stage 28 for immunohistochemistry with 12/101 antibody (d). a, b and c are representative phenotypes and f and g are close up views of d and e respectively. Arrowheads indicate regularly segmented somites on the injected side. (B) 20 ng of zfp36 morpholinos (MO) were injected into one ventral blastomere of 8-cell stage embryos with 250 pg of lacZ mRNA. Embryos were fixed at stage 29/30 (a, b, e, f), 33/34 (c, d) or 27 (g, h) and analysed for the expression of lim1, pax8, wnt4 and wt1 by in situ hybridization. |
