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Fig. 1.
Histone methyltransferase PRDM12 is expressed in the lateral pre-placodal ectoderm and regulated by Wnt and BMP signaling. (A) Stage RT-PCR analysis showing that PRDM12 mRNA is zygotically expressed after stage 13. Chordin and ODC were used as internal controls. (B–H) WISH analysis of PRDM12 (B, C and H), Six1 (D), and Foxd3 (F) expression at the indicated embryo stages. (E and G) Double in situ hybridization revealed partial co-localization of the PRDM12 transcripts (dark blue) with Six1 transcripts (light blue; E), but not with Foxd3 transcripts (light blue; G). The PRDM12 expression region is indicated by a red dotted outline in E and G. (H–S) Effects of BMP and Wnt signaling on PRDM12 and Pax3 expression at the neurula stage were analyzed by injecting Chordin mRNA (100 pg/embryo), BMP4 mRNA (20 pg/embryo), pCS2-Wnt3a (50 pg/embryo), and Dkk1 mRNA (10 pg/embryo) into the animal pole at the two-cell stage. (H and I) Dorsal view of the control embryo. (J–M) PRDM12 and Pax3 transcripts were expressed ectopically on the ventral side in embryos injected with Chordin mRNA (M, 76%, n=33). (N and O) BMP4 mRNA reduced PRDM12 and Pax3 expression (O, 100%, n=24). (P and Q) Posteriorized embryos caused the defective expression in the anterior part (Q, 91%, n=33). (R and S) Anteriorized embryos shifted the expression to the posterior region (S, 68%, n=31). YP; Yolk Plug. Scale bars represent 0.5 mm. |
