Dickinson K et al. (2006), Genomic profiling of mixer and Sox17beta target...

XB-ART-1118

Dev Dyn 2006 Feb 01;2352:368-81. doi: 10.1002/dvdy.20636.

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Genomic profiling of mixer and Sox17beta targets during Xenopus endoderm development.

Dickinson K , Leonard J , Baker JC .

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The transcription factors Mixer and Sox17beta have well-characterized roles in endoderm specification during Xenopus embryogenesis. In order to more thoroughly understand the mechanisms by which these endodermal regulators act, we expressed Mixer and Sox17beta in naïve ectodermal tissue and, using oligonucleotide-based microarrays, compared their genomic transcriptional profile to that of unaffected tissue. Using this approach, we identified 71 transcripts that are upregulated by Mixer or Sox17beta, 63 of which have previously uncharacterized roles in endoderm development. Furthermore, an in situ hybridization screen using antisense probes for several of these clones identified six targets of Mixer and/or Sox17beta that are expressed in the endoderm during gastrula stages, providing new and regional markers of the endoderm. Our results contribute further insight into the functions of Mixer and Sox17beta and bring us closer to understanding at the molecular level the pathways that regulate endoderm development.

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Species referenced: Xenopus laevis
Genes referenced: ascl1 btbd6 btg4 cdc42ep4 clcnkb cldn5 cmtm8 cpeb1 cxcr4 dmrta2 elf2 elovl2 eomes fgfr2 foxa1 foxc1 foxd1 gata5 gpr4 gpr4l.2 gsc hbg1 hprt1 irf1 irx4 itln1 itln2 kcnk6 lrsam1 mab21l2 marchf8 mig30 mixer msx2 myc nr2f2 odc1 otx2 pcdh10 pdgfra pisdl plk2 psmg2 rasl11b retreg2 sgk1 smad2 sox17a sox17b.1 sox17b.2 sp8 tbxt trabd2b traf4 tubg1 twist1 txnl4b usp33 vegt wnt11b XB22041735 XB5814120 XB5882685

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References [+] :

Afouda, GATA4, 5 and 6 mediate TGFbeta maintenance of endodermal gene expression in Xenopus embryos. 2005, Pubmed, Xenbase

	Figure 2. In situ hybridization screen reveals new expression patterns. A: Embryos at gastrula (10.5), neurula (15), and tailbud (30) stage are stained with antisense (left) and sense (right) probes for EST-11 showing expression in the mesoderm, neural tube, neural crest, and pronephros. B: Embryos at neurula and tailbud stage are stained with antisense and sense probes for EST-10 demonstrating expression in cement gland, neural tube nasal placode, otic placod, and forebrain. C: Embryos at gastrula and tailbud stage are stained for antisense and sense probes for EST-14 demonstrating expression in mesoderm and in a single posterior somite. D: Embryos at neurula and tailbud stage are stained with antisense and sense probes for EST-8, indicating expression within forebrain and heart. E: Embryos at neurula and tailbud stage are stained with antisense and sense probes for EST-34, showing expression in the placodes. F: Embryos at tailbud stage are stained with antisense and sense probes for EST-28 showing expression in nasal placodes and hindbrain. G: Embryos at neurula stage are stained with antisense and sense probes for EST-19, indicating expression within the neural tube. H: Embryos at tailbud stage are stained with antisense and sense probes for EST-9, demonstrating expression within pronephros and neural crest.
	Figure 3. Six transcripts are expressed in the early endoderm. The first two columns display stage 10.5 embryos, vegetal view, stained with antisense and sense probes for Cxcr4, March8, Borg4, EST-21, Gpr-4, and Xtwik-2. The last two columns display hemisected stage 10.5 embryos, lateral view, stained with antisense and sense probes for the same transcripts above. The descriptions along the right side indicate which transcripts were upregulated with either Mixer, Sox17 beta , or both. The arrow points to the deeper cells adjacent to the blastopore ring expressing March8.
	Figure 4. EST-21 and Gpr-4 have additional patterns during later embryonic stages. A,B: Embryos at tailbud stage are stained with antisense and sense probes for EST-21. C,D: Neurula stage embryos are stained with antisense and sense probes for Gpr-4. E,F: Tailbud stage embryos are stained with antisense and sense probes for Gpr-4.
	Fig. 2. Sp8 expression at stg. 30
	Fig. 4. Grp-4 expression at tailbud stg.
	fig. 2. Pcdh10 expression at stg 22-44
	Fig. 2. Btg4-a expression at tailbud stage
	Fig. 5. RT-PCR analysis confirms array data and supports an endogenous role for six new molecules in the endoderm pathway. RT-PCR was performed on cDNA synthesized from ectoderm explants expressing Mixer, Sox17 , Smad2, or VegT with primers for Cxcr4, March8, Borg4, EST-21, Gpr-4, and Xtwik-2. -gal was injected as a control. Primers for Sox17 and Xbra were used as positive controls. ODC was used as a loading control. WE, whole embryo; RT, minus reverse transcriptase.

	loc100492494 (fish-egg lectin-like) gene expression in bissected Xenopus laevis embryo, assayed via in situ hybridization, NF stage 10.5, horizontal view.
	loc100492494 (fish-egg lectin-like) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anterior left, dorsal up.
	plk2 (polo-like kinase 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.