Haremaki T , Sridharan J , Dvora S , Weinstein DC .

R01-GM61671 NIGMS NIH HHS , R01 GM061671-09 NIGMS NIH HHS , R01 GM061671 NIGMS NIH HHS

	Figure 2. The Eif4a3 morphant phenotype. A: Loss of pigment-forming cells following Eif4a3 knockdown. Twenty-one nanograms of each morpholino (“4a3MO”: Eif4a3 morpholino; “5MM”: 5 base pair mismatch Eif4a3 morpholino) was injected, as listed, in this and subsequent figures. “Rescue”: Eif4a3MO+2 ng eif4a3 RNA. Embryos were derived from albino eggs fertilized with sperm from wild-type males. B: Loss of touch-response behavior in Eif4a3 morphants. Graph depicting percentage of touch-responsive embryos injected with listed doses of eif4a3 RNA and/or morpholino (4a3MO), as shown. C: Defects of heart formation in Eif4a3 morphants. Troponin T staining of cardiac tissue in stage-37/38 embryos injected with 4a3MO, 5MM, or 4a3MO+Eif4a3 RNA (Rescue). Note lack of heart looping in 4a3MO-injected embryo. D: Whole-mount in situ hybridization with antisense probes against Xnkx-2.5 in stage-28 embryos injected with Eif4a3MO (4a3MO) or Eif4a3MM (5MM), as listed. No significant differences were seen in the expression of this marker at neurula or tailbud stages (data not shown).
	Figure 3. The Y14 and Magoh morphant phenotypes. Loss of pigment-forming cells following Y14 or Magoh knockdown. Y14 morpholino (2.6 ng) (“Y14MO”) or 10 ng of Magoh morpholino (“MagohMO”) was injected, as listed. “Rescue”: MO+2 ng corresponding RNA. Embryos were derived from albino eggs fertilized with sperm from wild-type males.
	Figure 5. Somite and neuronal development in Eif4a3 morphants. A: 12/101 antibody staining of somitic mesoderm in embryos injected with Eif4a3MO (4a3MO) or a control (scrambled) morpholino (CMO). No differences in somite staining were observed between the two populations. B: Whole-mount in situ hybridization with antisense probes against elrC (left) or sox3 (right) in embryos injected with Eif4a3MO (4a3MO) or Eif4a3MM (5MM). Note slight expansion of anterior neural tube in Eif4a3 morphants. C: Left panels: 3A10 antibody staining of neurons in embryos injected with Eif4a3MO (4a3MO) or a control (scrambled) morpholino (CMO). Right panels: High magnification of trunk detail of embryos at left. Note disorganization of “Y”-shaped motor neuron bundles in Eif4a3 morphants.
	Figure 6. Eif4a3 morphants display late defects in sensory neuron and neural crest development. A: Islet-1 antibody staining of Rohon-Beard (top band of nuclei in top panel, arrows) and motor (bottom band of nuclei in top panel, arrowheads) neurons. Increased dorsoventral spread of Islet-1-positive Rohon-Beard cells, as well as a reduction in intensity of Islet-1 staining in motor neuron nuclei, was observed in Eif4a3 morphants; both defects were partially rescued by co-injection of eif4a3 RNA. B: Whole-mount in situ hybridization with an antisense probe against hox11L2 in embryos injected with Eif4a3MO (4a3MO) or a control (scrambled) morpholino (CMO). Note disorganization of hox11L2-positive Rohon-Beard cells (arrows) in the Eif4a3 morphant embryo. C–F: Whole-mount in situ hybridization with antisense probes against slug/snail2 (C), sox10 (D), Xtwist (E), or Xsix-1 (F) in embryos injected with Eif4a3MO (4a3MO) or Eif4a3MM (5MM) morpholinos, as listed. No consistent differences were seen in the expression of these markers at neurula or tailbud stages.
	Figure 7. Eif4a3 is required for expression of tyrosinase genes. Whole-mount in situ hybridization with antisense probes against tyrosinase (tyr) (top panels) or tyrosinase-related protein-2 (dct) (bottom panels) in embryos injected with Eif4a3MO (4a3MO), Eif4a3MM (5MM), or Eif4a3MO+2 ng eif4a3 RNA (Rescue). With both probes, intensity of signal and number of positive cells were reduced in Eif4a3 morphants at tailbud stages; this effect was rescued by co-expression of eif4a3 RNA.
	Figure 8. Apoptotic cell death is increased in Eif4a3 morphants. A: TUNEL assay of embryos injected with Eif4a3MO (4a3MO), Eif4a3MM (5MM), or Eif4a3MO+2 ng eif4a3 RNA (Rescue) at stage 32. Strong staining was observed in head only. B: Immunohistochemistry using Caspase 3 antibodies in embryos injected with Eif4a3MO (4a3MO), Eif4a3MM (5MM), or Eif4a3MO+2 ng eif4a3 RNA (Rescue); signal was observed in the eyes of Eif4a3 morphants. C: Phosphohistone H3 antibody staining (blue) of embryos injected on one side (arrowhead) with Eif4a3MO (4a3MO) and 500 pg β-gal RNA as a lineage trace (red) at stage 32. Phosphohistone H3 signal was not significantly affected by Eif4a3 knockdown.

Artinger, Zebrafish narrowminded suggests a genetic link between formation of neural crest and primary sensory neurons. 1999, Pubmed

Artinger, Zebrafish narrowminded suggests a genetic link between formation of neural crest and primary sensory neurons. 1999, Pubmed
Ballut, The exon junction core complex is locked onto RNA by inhibition of eIF4AIII ATPase activity. 2005, Pubmed
Chang, The nonsense-mediated decay RNA surveillance pathway. 2007, Pubmed
Chen, Maintenance of motor neuron progenitors in Xenopus requires a novel localized cyclin. 2007, Pubmed , Xenbase
Clarke, Sensory physiology, anatomy and immunohistochemistry of Rohon-Beard neurones in embryos of Xenopus laevis. 1984, Pubmed , Xenbase
Cornell, Delta/Notch signaling promotes formation of zebrafish neural crest by repressing Neurogenin 1 function. 2002, Pubmed
Cornell, Delta signaling mediates segregation of neural crest and spinal sensory neurons from zebrafish lateral neural plate. 2000, Pubmed
Ericson, Two critical periods of Sonic Hedgehog signaling required for the specification of motor neuron identity. 1996, Pubmed
Gingras, eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation. 1999, Pubmed
Giorgi, The EJC factor eIF4AIII modulates synaptic strength and neuronal protein expression. 2007, Pubmed
Hemmati-Brivanlou, Inhibition of activin receptor signaling promotes neuralization in Xenopus. 1994, Pubmed , Xenbase
Hensey, Programmed cell death during Xenopus development: a spatio-temporal analysis. 1998, Pubmed , Xenbase
Hopwood, A Xenopus mRNA related to Drosophila twist is expressed in response to induction in the mesoderm and the neural crest. 1989, Pubmed , Xenbase
Kenwrick, Pilot morpholino screen in Xenopus tropicalis identifies a novel gene involved in head development. 2004, Pubmed , Xenbase
Khajavi, Nonsense-mediated mRNA decay modulates clinical outcome of genetic disease. 2006, Pubmed
Kolker, Confocal imaging of early heart development in Xenopus laevis. 2000, Pubmed , Xenbase
Kumasaka, Isolation and developmental expression of tyrosinase family genes in Xenopus laevis. 2003, Pubmed , Xenbase
Le Hir, The spliceosome deposits multiple proteins 20-24 nucleotides upstream of mRNA exon-exon junctions. 2000, Pubmed , Xenbase
Li, Eukaryotic translation initiation factor 4AIII (eIF4AIII) is functionally distinct from eIF4AI and eIF4AII. 1999, Pubmed , Xenbase
Linker, Relationship between gene expression domains of Xsnail, Xslug, and Xtwist and cell movement in the prospective neural crest of Xenopus. 2000, Pubmed , Xenbase
Martinsen, Cardiac neural crest ablation alters Id2 gene expression in the developing heart. 2004, Pubmed , Xenbase
Moore, Pre-mRNA processing reaches back to transcription and ahead to translation. 2009, Pubmed
Owttrim, Divergent genes for translation initiation factor eIF-4A are coordinately expressed in tobacco. 1991, Pubmed
Pandur, Xenopus Six1 gene is expressed in neurogenic cranial placodes and maintained in the differentiating lateral lines. 2000, Pubmed , Xenbase
Patterson, Hox11-family genes XHox11 and XHox11L2 in xenopus: XHox11L2 expression is restricted to a subset of the primary sensory neurons. 1999, Pubmed , Xenbase
Roberts, Early functional organization of spinal neurons in developing lower vertebrates. 2000, Pubmed , Xenbase
Saka, Spatial and temporal patterns of cell division during early Xenopus embryogenesis. 2001, Pubmed , Xenbase
Schlosser, Induction and specification of cranial placodes. 2006, Pubmed , Xenbase
Shibuya, eIF4AIII binds spliced mRNA in the exon junction complex and is essential for nonsense-mediated decay. 2004, Pubmed
Steventon, Differential requirements of BMP and Wnt signalling during gastrulation and neurulation define two steps in neural crest induction. 2009, Pubmed , Xenbase
Suri, Xema, a foxi-class gene expressed in the gastrula stage Xenopus ectoderm, is required for the suppression of mesendoderm. 2005, Pubmed , Xenbase
Tange, The ever-increasing complexities of the exon junction complex. 2004, Pubmed
Tonissen, XNkx-2.5, a Xenopus gene related to Nkx-2.5 and tinman: evidence for a conserved role in cardiac development. 1994, Pubmed , Xenbase
Weinstein, Epidermal induction and inhibition of neural fate by translation initiation factor 4AIII. 1997, Pubmed , Xenbase
Wilson, Induction of epidermis and inhibition of neural fate by Bmp-4. 1995, Pubmed , Xenbase