Hong CS , Saint-Jeannet JP .

R01 DE014212 NIDCR NIH HHS, R01 DE025468 NIDCR NIH HHS

             rhombomere morphogenesis
             hindbrain development

	Figure 2. Developmental expression of znf703. (a–d) By in situ hybridization at the neurula stage (NF stage 13/14) znf703 is detected in the dorso-lateral ectoderm posteriorly, including the neural plate and the neural crest forming region, with a gap of expression at the dorsal midline. (e and f) At stages 16 through 19, znf703 expression persists in the dorsal ectoderm where it appears to be confined to the prospective hindbrain and spinal cord. (g) Transverse section of a stage 16 embryo, dorsal to top, showing znf703 expression in the neural plate (np), neural crest (nc), ventral non-neural ectoderm (nne) and lateral plate mesoderm (lpm). (k–m) Later in development znf703 is diffusely expressed throughout the ectoderm, and enriched in the hatching gland (hg) and the branchial arches (ba). (a, c, e, h, and k) dorsal views, anterior to top. (b, d, j, l, and m) lateral views, dorsal to top, anterior to right. (f, and i) frontal views, dorsal to top. The embryonic stages are indicated in the lower left corner of each panel
	Figure 3. znf703 anterior expression domain corresponds to r3/r4. (a) At stage 15, znf703 is detected dorsally, with a sharp anterior boundary of expression. (b–d) Double in situ hybridization showing the spatial relationship of znf703 with snai2 in the neural crest (b), mafb in r5/r6 (c), and fgf3 in r3/r4 (d) Dorsal views, anterior to top. The yellow lines indicate the position of the most anterior boundary of znf703 expression. The red asterisks indication the position of the most anterior expression domains of snai2, mafb, and fgf3. Insets (lower right corner) in panels (b), (e), and (d) depict single in situ hybridization for snai2, mafb, and fgf3, respectively
	Figure 4. znf703 is a true target of Pax3 and Zic1. (a and b) Fold induction of snai2 (a) and znf703 (b) by expression of pax3GR and zic1GR in animal cap explants (microarray data; Bae et al., 2014). (c and d) Injection in one blastomere at the two-cell stage of MOs blocking Pax3 (pax3MO; 40 ng) Zic1 (zic1MO; 40 ng) function resulted in a strong reduction of snai2 expression (c),r while only the most anterior expression domain of znf703 was affected (d) at the neurula stage. The injected side is to the right as indicated by the presence of the lineage tracer (Red-Gal). The percentage of affected embryos is indicated in the upper right corner. Dorsal views, anterior to top.
	Figure 5. Regulation of znf703 expression by neural crest inducing signals. (a) mRNAs encoding noggin (1 ng) and wnt1 (100 pg) were injected, alone or in combination with pax3MO, into both blastomeres in the animal pole region at the 2-cell stage. At the blastula stage (NF Stage 9), animal cap explants were dissected, cultured for 8 hr at room temperature (equivalent Stage 13/14) and analyzed by qRT-PCR. (b) In combination, Noggin and Wnt1 induce snai2 and repress keratin expression. Co-injection of pax3MO blocks snai2 induction by Noggin and Wnt1 and promotes neural fate (sox2 expression). The induction of znf703 by Noggin and Wnt1 is unaffected by injection of pax3MO
	Figure 6. Znf703 is required for posterior hindbrain patterning. (a) The target sequence of Znf703 morpholino antisense oligonucleotide (Znf703MO; grey box) is indicated. The position of the start codon (ATG) is indicated. (b) In vitro coupled transcription/translation reactions with plasmid encoding Znf703. Increasing amounts of Znf703MO, 10 ng (+), 100 ng (++) and 1000 ng (+++), block translation directed by znf703 mRNA. (c–j) Unilateral injection of znf703MO (45 ng) at the two-cell stage expanded egr4 (c), hoxb1 (d) and fgf3 (e) expression domains in the hindbrain at the neurula stage (NF Stage 14/15). In contrast mafb (f) and the r5 expression domain of egr2 were reduced or lost (g). In morphant embryos the expression domain of sox2 was expanded (h), while the neural crest markers were mildly (snai2; g) or strongly (sox10; h) reduced. The horizontal lines (f) indicate the width of the neural plate on the control (black line) and injected (white line) sides. The injected side is to the right as indicated by the presence of the lineage tracer (Red-Gal). The percentage of affected embryos is indicated in the upper right corner. Dorsal views, anterior to top
	Figure 7. Znf703 overexpression affects hindbrain and neural crest development. (a–h) Unilateral injection of znf703 mRNA (1 ng) at the two-cell stage significantly reduced the expression domain of egr4 (a), hoxb1 (b), fgf3 (c) mafb (d), and egr2 (e), in the hindbrain at the neurula stage (NF Stage 14/15). In these embryos sox2 expression domain was expanded (f), and both neural crest markers, snai2 (g) and sox10 (h), were strongly reduced. The horizontal lines (f) indicate the width of the neural plate on the control (black line) and injected (white line) sides. The injected side is to the right as indicated by the presence of the lineage tracer (Red-Gal). The percentage of affected embryos is indicated in the upper right corner. Dorsal views, anterior to top
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 13, dorsal view, anterior up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 13, lateral view, anterior right, dorsal up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 14, lateral view, anterior right, dorsal up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 16, (e) dorsal view, anterior up and (f) anterior view,, dorsal up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19 (i), anterior view, dorsal up and (j) lateral view, anterior right, dorsal up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25, dorsal view, anterior up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25, anterior/lateral view, dorsal up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25, lateral view, anterior right, dorsal up.
	znf703 (zinc finger protein 703) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 16, transverse section, dorsal up, showing expression in the non-neural ectoderm (nne) and the underlying lateral plate mesoderm (lpm), as well as neural crest ( nc) and neural plate (np)

Aoki, Sox10 regulates the development of neural crest-derived melanocytes in Xenopus. 2003, Pubmed, Xenbase

Aoki, Sox10 regulates the development of neural crest-derived melanocytes in Xenopus. 2003, Pubmed , Xenbase
Bae, A novel function for Egr4 in posterior hindbrain development. 2015, Pubmed , Xenbase
Bae, Identification of Pax3 and Zic1 targets in the developing neural crest. 2014, Pubmed , Xenbase
Betancur, Assembling neural crest regulatory circuits into a gene regulatory network. 2010, Pubmed
Bradley, The structure and expression of the Xenopus Krox-20 gene: conserved and divergent patterns of expression in rhombomeres and neural crest. 1993, Pubmed , Xenbase
Dorfman, Elbow and Noc define a family of zinc finger proteins controlling morphogenesis of specific tracheal branches. 2002, Pubmed
Garnett, BMP, Wnt and FGF signals are integrated through evolutionarily conserved enhancers to achieve robust expression of Pax3 and Zic genes at the zebrafish neural plate border. 2012, Pubmed
Godsave, Expression patterns of Hoxb genes in the Xenopus embryo suggest roles in anteroposterior specification of the hindbrain and in dorsoventral patterning of the mesoderm. 1994, Pubmed , Xenbase
Gutkovich, Xenopus Meis3 protein lies at a nexus downstream to Zic1 and Pax3 proteins, regulating multiple cell-fates during early nervous system development. 2010, Pubmed , Xenbase
Harland, In situ hybridization: an improved whole-mount method for Xenopus embryos. 1991, Pubmed , Xenbase
Holland, ZNF703 is a common Luminal B breast cancer oncogene that differentially regulates luminal and basal progenitors in human mammary epithelium. 2011, Pubmed
Hong, The activity of Pax3 and Zic1 regulates three distinct cell fates at the neural plate border. 2007, Pubmed , Xenbase
Hong, Transcription factor AP2 epsilon (Tfap2e) regulates neural crest specification in Xenopus. 2014, Pubmed , Xenbase
Hoyle, nlz gene family is required for hindbrain patterning in the zebrafish. 2004, Pubmed
Huang, Induction of the neural crest and the opportunities of life on the edge. 2004, Pubmed , Xenbase
Ishibashi, Distinct roles of maf genes during Xenopus lens development. 2001, Pubmed , Xenbase
Ji, Nolz1 is induced by retinoid signals and controls motoneuron subtype identity through distinct repressor activities. 2009, Pubmed
Kjolby, Genome-wide identification of Wnt/β-catenin transcriptional targets during Xenopus gastrulation. 2017, Pubmed , Xenbase
Kolm, Efficient hormone-inducible protein function in Xenopus laevis. 1995, Pubmed , Xenbase
LaBonne, Snail-related transcriptional repressors are required in Xenopus for both the induction of the neural crest and its subsequent migration. 2000, Pubmed , Xenbase
Li, Conserved gene regulatory module specifies lateral neural borders across bilaterians. 2017, Pubmed , Xenbase
Lombardo, Expression and functions of FGF-3 in Xenopus development. 1998, Pubmed , Xenbase
Mayor, Induction of the prospective neural crest of Xenopus. 1995, Pubmed , Xenbase
McGlinn, Expression of the NET family member Zfp503 is regulated by hedgehog and BMP signaling in the limb. 2008, Pubmed
Milet, Pax3 and Zic1 drive induction and differentiation of multipotent, migratory, and functional neural crest in Xenopus embryos. 2013, Pubmed , Xenbase
Mizuseki, Xenopus Zic-related-1 and Sox-2, two factors induced by chordin, have distinct activities in the initiation of neural induction. 1998, Pubmed , Xenbase
Monsoro-Burq, Msx1 and Pax3 cooperate to mediate FGF8 and WNT signals during Xenopus neural crest induction. 2005, Pubmed , Xenbase
Nakamura, Nlz1/Znf703 acts as a repressor of transcription. 2008, Pubmed
Nakamura, A novel subfamily of zinc finger genes involved in embryonic development. 2004, Pubmed
Pereira, Evolution of the NET (NocA, Nlz, Elbow, TLP-1) protein family in metazoans: insights from expression data and phylogenetic analysis. 2016, Pubmed
Plouhinec, Pax3 and Zic1 trigger the early neural crest gene regulatory network by the direct activation of multiple key neural crest specifiers. 2014, Pubmed , Xenbase
Runko, Nlz belongs to a family of zinc-finger-containing repressors and controls segmental gene expression in the zebrafish hindbrain. 2003, Pubmed
Sadaghiani, Neural crest development in the Xenopus laevis embryo, studied by interspecific transplantation and scanning electron microscopy. 1987, Pubmed , Xenbase
Saint-Jeannet, Regulation of dorsal fate in the neuraxis by Wnt-1 and Wnt-3a. 1997, Pubmed , Xenbase
Santagati, Cranial neural crest and the building of the vertebrate head. 2003, Pubmed
Sato, Neural crest determination by co-activation of Pax3 and Zic1 genes in Xenopus ectoderm. 2005, Pubmed , Xenbase
Sauka-Spengler, A gene regulatory network orchestrates neural crest formation. 2008, Pubmed
Simões-Costa, Establishing neural crest identity: a gene regulatory recipe. 2015, Pubmed
Sircoulomb, ZNF703 gene amplification at 8p12 specifies luminal B breast cancer. 2011, Pubmed
Slack, An interaction between dorsal and ventral regions of the marginal zone in early amphibian embryos. 1980, Pubmed , Xenbase
Smith, Expression cloning of noggin, a new dorsalizing factor localized to the Spemann organizer in Xenopus embryos. 1992, Pubmed , Xenbase
Stuhlmiller, Current perspectives of the signaling pathways directing neural crest induction. 2012, Pubmed , Xenbase
Trainor, Patterning the cranial neural crest: hindbrain segmentation and Hox gene plasticity. 2000, Pubmed
Wolda, Overlapping expression of Xwnt-3A and Xwnt-1 in neural tissue of Xenopus laevis embryos. 1993, Pubmed , Xenbase