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Experiment details for hoxb3

Dibner C et al. (2001) Assay

XMeis3 protein activity is required for proper hindbrain patterning in Xenopus laevis embryos.

Gene Clone Species Stages Anatomy
hoxb3.S laevis NF stage 21 hindbrain , neuroectoderm , rhombomere R5 , rhombomere R6

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  Fig. 3. Expression pattern of neural markers in embryos injected with XMeis3-AM RNA. Two-cell albino embryos were injected unilaterally into the animal hemisphere of one blastomere with 50-100 pg of XMeis3−AM RNA. The red arrow delineates the dorsal midline. In all embryos, XMeis3-AM injection is on the left side. In all cases (except Fig. 3M), embryos are viewed dorsally; embryos are oriented anterior (top) to posterior (bottom). (A) In situ hybridization with otx2. Expression is expanded posteriorly on the XMeis3-AM injected side. The red lines delineate the AP extent of otx2 expression on the uninjected versus injected side. (B) In situ hybridization with otx2 and Krox20. otx2 expression is expanded posteriorly and Krox20 expression (blue arrows) is lost on the XMeis3-AM-injected side. The red lines delineate the AP extent of otx2 expression on the uninjected versus injected side. (C) In situ hybridization with otx2 and En2 (red). Expression of otx2 and En2 (blue arrows) is expanded posteriorly. (D) In situ hybridization with cpl-1 and En2 (red). cpl-1 expression is expanded posteriorly and En2 expression (blue arrow/uninjected side) is lost on the XMeis3-AM-injected side. The red lines delineate the AP extent of cpl-1 expression on the uninjected versus injected side. (E) In situ hybridization with XAG1; expression is expanded posteriorly and laterally on the XMeis3-AM injected side. (F) In situ hybridization with nrp1; expression is unchanged on the XMeis3-AM injected side. (G) In situ hybridization with Krox20 and HoxB9; Krox20 expression (blue arrows/uninjected side) is eliminated on the XMeis3-AM-injected side. HoxB9 expression is unchanged on the XMeis3-AM injected side. (H) In situ hybridization with XE10. XE10 expression is eliminated on the XMeis3-AM-injected side. (I) In situ hybridization with HoxB1. HoxB1 expression is eliminated on the XMeis3-AM injected side. (J) In situ hybridization with HoxB3. HoxB3 expression is eliminated on the XMeis3-AM-injected side. (K) In situ hybridization with En2 (red) and Krox20. En2 expression is pushed posteriorly to the r3/r4 boarder and Krox20 expression is pushed posteriorly to the r5/r7 boarder on the XMeis3-AM-injected side. (L) In situ hybridization with En2 and Krox20. En2 expression is pushed posteriorly to r3 on the XMeis3-AM-injected side. The blue arrow delineates the reduced Krox20 expression (fused stripes) on the injected side. (M) In situ hybridization with En2 (red) and Krox20. Expression of En2 and Krox20 (blue arrows/uninjected side) is eliminated on the XMeis3-AM-injected side. An anterior view of the embryo: dorsal (top) to ventral (bottom). (N) In situ hybridization with n-tubulin. n-tubulin expression is eliminated on the XMeis3-AM-injected side. The trigeminal neuron is marked by blue arrows on both sides.

Gene Clone Species Stages Anatomy
hoxb3.S laevis NF stage 21 hindbrain , neuroectoderm , rhombomere R5 , rhombomere R6

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  Fig. 6. XMeis3 antisense morpholino oligonucleotide eliminates hindbrain marker expression. Two-cell albino embryos were injected unilaterally into the animal hemisphere of one blastomere with 6-7.5 ng of the XMeis3 AMO. In situ hybridization was performed in late neurula stage embryos. In all cases, embryos are viewed dorsally; embryos are oriented anterior (top) to posterior (bottom). The red arrow delineates the dorsal midline. Embryos were injected on the right side. (A) In situ hybridization with Krox20 and HoxB9; Krox20 expression is eliminated on the AMO-injected side. HoxB9 expression is unchanged on the AM- injected side. (B) In situ hybridization with XE10; expression is eliminated on the AMO-injected side. (C) In situ hybridization with HoxB3 and HoxB9; HoxB3 expression is eliminated on the AMO-injected side. HoxB9 expression is unchanged on the AMO-injected side. (D) In situ hybridization with XMeis3 and En2 (red); expression of both markers is posteriorized on the AMO-injected side. (E) In situ hybridization with XMeis3; expression is inhibited on the AMO-injected side. The XMeis3 expression in r2 is indicated by arrows on both sides.