Faucheux C et al. (2010), Vestigial like gene family expression in Xenopu...

XB-ART-41925

Int J Dev Biol 2010 Jan 01;548-9:1375-82. doi: 10.1387/ijdb.103080cf.

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Vestigial like gene family expression in Xenopus: common and divergent features with other vertebrates.

Faucheux C , Naye F .

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The Drosophila Vestigial and Scalloped proteins form heterodimers that control wing development and are involved in muscle differentiation. Four vestigial like genes have been described in mammals. Similar to the Drosophila vestigial gene, they encode a short conserved domain (TONDU) required for interaction with the mammalian paralogues of Drosophila Scalloped (i.e., TEAD proteins). We previously identified two TEAD genes in Xenopus laevis and we report here the expression of four distinct vestigial like genes in Xenopus (vgll1-4) that represent amphibian orthologs of the mammalian vestigial like genes. Vgll1 has a unique expression pattern which is restricted to epidermal cells, both in the embryo and in the adult. Vgll2 is expressed in the skeletal muscle lineage downstream of myogenic factors and in the embryonic brain similar to the avian and mammalian orthologues. Vgll3 expression is transient, identifies embryonic hindbrain rhombomere 2, and is negatively regulated by en2, but not by egr2. Vgll4 is mainly expressed in anterior neural structures. In summary, the four Xenopus vgll genes have unique/complex expression profiles and they are differently expressed during embryogenesis. Moreover, these amphibian vestigial like genes display distinct responses to the major signaling pathways (i.e., activin, FGF or BMP) that orchestrate pattern-formation during early development.

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Species referenced: Xenopus laevis
Genes referenced: bmp4 egr2 en2 fgf2 hey1 msx1 myf5 myl1 myod1 odc1 tcf3 tead1 tead4 uqcc6 ventx1.2 vgll1 vgll2 vgll3 vgll4

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	Fig. 3. Spatial expression analysis of Xenopus laevis vestigial like genes in embryo by in situ hybridization. (A) vgll2 expression. (a) dorsal view; (b,c,e) lateral views; (d,f,g,h) transversal sections. Double in situ hybridization of stage 30 embryo with vgll2 and myod1 (g) or vgll2 and myl1 (h). (B) vgll3 expression. (a,b,f,g), frontal views; (c) dorsal view; (e) lateral view. (f) double in situ hybridization of stage 16 embryo with vgll3 and en2 (f) or vgll3 and egr2 (g). (d) parasagittal section of the embryo shown in g; (h) higher magnification of d. The position of rhombomeres is indicated (r2, r3 and r5). (C) vgll4 expression. (a) anterior view; (b) dorsal view; (c) lateral view; (d) transversal section. The positions of the section were indicated. Embryo staging according to Nieuwkoop and Faber (1967) is indicated. ba, branchial arches; br, brain; ey, eye; h, hypophyseal anlage; hm, head mesenchyme; hym, hypaxial muscle; lp, lens placode; m, mesencephalon; mc, mandibular neural crest; n, notochord; op, olfactory placode; ov, otic vesicle; r, rhombomere; s, somites.
	Fig. 4. Xenopus laevis vestigial like gene regulation. (A) Animal caps treated with different amounts of activin or bFGF (5, 25 or 100 ng/ml) were analyzed by RT-PCR for the expression of the different vgll genes. For compari- son the expression of fast skeletal myosin light chain (myl1) and epidermal keratin(krt) genes was assayed. (B) Animal cap explants from stage 9 embryos were dissoci- ated into deep layer (Inner) cells and superficial layer (Outer) and assayed for vgll1, vgll4, tead1 and tead2 genes expression. Expression of hey1 and epidermal keratin (krt) genes was assayed in control. RNA from intact caps is used as control (Cap). (C) Dissociated inner cells from animal caps were treated with 1μg/ml of BMP4 (BMP4) for one hour or not (-) and assayed for the expression of vgll1, epidermal keratin (krt) and vent1. (D) 250 pg of tBR4 mRNA were injected into 2-cell stage embryos and animal cap explants were assayed by RT- PCR for the expression of vgll1, epidermal keratin (krt) and msx1. (E) 2-cell stage embryos were injected with 1ng of myogenic factors encoding mRNA (myod1, myf5 or myf6) plus 1ng of tcf3 (formerly E12) mRNA or 1ng of vgll2 mRNA alone. Animal cap explants from injected embryos or uninjected embryos (-) were assayed by RT- PCR for the expression of vgll2, myod1 or myl1. In (A,C,D,E), analyses were performed when control em- bryo (Emb) reaches stage 20. (-) Odc was used as control of loading and a reaction was performed in absence of reverse transcriptase to check for genomic DNA con- tamination (-RT). (F-H) In situ analysis of vgll3 expression in neurula embryo (frontal view): F, control embryo; G, H embryo injected with the indicated mRNA (egr2, en2). LacZ mRNA was used as a tracer in G, H. Uninjected side (ni) was used as control.
	vgll2 (vegtigial like 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19, dorsal view, anterior left.
	vgll2 (vegtigial like 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anterior left, dorsal up.



	vgll3 (vegtigial like 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 15, anterior view, dorsal up.
	vgll3 (vegtigial like 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 16, sagittal section, anterior left, dorsal up.
	vgll3 (vegtigial like 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 18, anterior view, dorsal up.
	vgll3 (vegtigial like 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 24, dorsal view, anterior left.
	vgll4 (vegtigial like 4) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 16, anterior view, dorsal up.
	vgll4 (vegtigial like 4) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 23, dorsal view, anterior left, dorsal up.