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Figure 1 | Phenotypes of loss- and gain-of-function of head organizer TFs in frogs. (a) Expression patterns of otx2, otx5, lim1 and gsc in Xenopus gastrula embryos. Schematic representation shows co-expression of lim1, otx2, otx5 and gsc in the head organizer 49,16,22,23,56 and lim1 expression in the trunk organizers16,56. A double-headed arrow indicates the dorsal (D) and ventral (V) axis. Co-expression of these genes in X. tropicalis early gastrula embryos was shown with in situ hybridization using hemisections. In anterior neuroectoderm, otx2 is weakly expressed, while otx5 is more strongly expressed. Arrowhead, dorsal blastopore; open arrowhead, expression in the anterior neuroectoderm; and dashed line, boundary of ectoderm and mesoderm. (b) Loss- of-function analysis in X. tropicalis. Phenotypes of normal embryos (control morphants) and head-reduced embryos (lim1/otx2/otx5/gsc morphants) are shown. Sagittal section and whole mount in situ hybridization of the telencephalon marker (eomes) and the midbrainhindbrain boundary marker (en2) showed that forebrain, midbrain and foregut were shrunk in head-reduced embryos. Note that the notochord reached the anterior-most region in the morphant. Tailbud embryos are shown in lateral (anterior to the left; three left panels) or dorsoanterior (right-most panels) views. fb, forebrain; fg, foregut; no, notochord. (c) Gain-of-function analysis in X. laevis. Left panels show lateral views of a secondary head with one eye generated after ventral injection (inj.) of a cocktail of mRNAs as indicated. Right panels show immunostaining of head organizer cocktail-injected embryos against notochord by MZ15 (dorsal view of the same embryo as shown in the left panel) and somites by 12/101 (lateral view), respectively. Scale bar, 250 mm.

Image published in: Yasuoka Y et al. (2014)

Copyright © 2014. Image reproduced with permission of the Publisher, Macmillan Publishers Ltd.

GeneSynonymsSpeciesStage(s)Tissue
lhx1.Llim-1, Lim1, lim1, Xlim-1, Xlim1X. laevisThroughout NF stage 10.5dorsal marginal zone
upper blastopore lip
dorsal involuting marginal zone
otx2.Lotx-2, otx2-a, otx2-b, otxA, Xotx-2, Xotx2X. laevisThroughout NF stage 10.5dorsal marginal zone
upper blastopore lip
dorsal involuting marginal zone
crx.Lcrx-a, crx-b, otx5, otx5b, Xotx5, Xotx5bX. laevisThroughout NF stage 12involuted dorsal mesoderm
presumptive axial mesoderm
gsc.Lgoosecoid, gsc-a, gsc-b, XgscX. laevisThroughout NF stage 12involuted dorsal mesoderm
presumptive axial mesoderm
en2.LEn-2, en2-a, en2-b, eng2, engrailed 2, engrailed homolog 2, engrailed-2, maben, xen2X. laevisSometime during NF stage 31 to NF stage 32midbrain-hindbrain boundary
eomes.Leomesodermin, tbr2, xeomesX. laevisSometime during NF stage 31 to NF stage 32forebrain

Image source: Published

Experiment + Assay Source Phenotypes and Disease
Xtr Wt + otx2 MO + crx MO + gsc MO + lhx1 MO + NF31-32 (whole-mount microscopy) Fig. 1 b row2
Anatomical Phenotype
abnormal forebrain
abnormal foregut morphology
abnormal head morphology
absent eye
absent optic stalk
decreased size of the forebrain
decreased size of the midbrain-hindbrain boundary
Expression Phenotype
absent eomes expression in optic stalk
Xla Wt + otx2 + ssbp3 + ldb1 + lhx1 + NF37/38-39 (whole-mount microscopy) Fig. 1. C
Anatomical Phenotype
duplicated eye
duplicated head
duplicated main body axis
Xla Wt + tle1 + gsc + otx2 + ssbp3 + ldb1 + lhx1 mRNA + NF37/38-39 (whole-mount microscopy) Fig. 1. C
Anatomical Phenotype
duplicated eye
duplicated head
duplicated main body axis

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