Cyclin D1 and Her9 control the proliferation of boundary cells. (A-D) Tg[BCP:GFP] embryos at the indicated stages were in situ hybridized with ccnd1 and cdkn1ca or her9 probes. (E,F) Tg[BCP:H2BGFP;ccnd1+/+] and Tg[BCP:H2BGFP;ccnd1pfu1/pfu1] embryos displaying boundary cells in green at 26?hpf . (G) Plot showing the number of boundary cells (87.3±16.5 in ccnd1+/+ n=4 versus 61.3±10.8 in ccnd1pfu1/pfu1 n=8; *P=0.04, Welch's test). (H-K) Tg[BCP:H2BGFP;ccnd1+/+] and Tg[BCP:H2BGFP;ccnd1pfu1/pfu1] embryos injected with control-MO (H,I) or her9-MO (J,K). (L) Plot showing the number of boundary cells in Tg[BCP:H2BGFP;ccnd1+/+] and Tg[BCP:H2BGFP;ccnd1pfu1/pfu1] embryos upon different conditions. (H) 75.7±15.9 in ccnd1+/+ control-MO n=3; (I) 44.7±19.4 in ccnd1pfu1/pfu1 control-MO n=3; (J) 46.8±7.7 in ccnd1+/+ her9-MO n=6; (K) 46.5±16.1 in ccnd1pfu1/pfu1 her9-MO n=6. H versus I, *P=0.04; H versus J, *P=0.03; H versus K, *P=0.03; I versus J, P=0.99; I versus K, P=0.99; J versus K, P>0.99. One-way ANOVA, Dunnett's multiple comparison test. Images are transverse views of the r4/r5 boundary (E,F,H-K,a-d) or dorsal maximum intensity projections of the hindbrain with anterior to the left (A-D). The plots show mean±s.d. Dotted lines delimitate the contour of the neural tube. Arrowheads indicate the position of the hindbrain boundaries. BCP, boundary cell population; hpf, hours post-fertilization; MO, morpholino; ns, not significant. Scale bars: 50?µm.
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