Zhang Q et al. (1999), Vg1 RBP intracellular distribution and evolutio...

XB-ART-12131

Mech Dev 1999 Oct 01;881:101-6. doi: 10.1016/s0925-4773(99)00162-8.

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Vg1 RBP intracellular distribution and evolutionarily conserved expression at multiple stages during development.

Zhang Q , Yaniv K , Oberman F , Wolke U , Git A , Fromer M , Taylor WL , Meyer D , Standart N , Raz E , Yisraeli JK .

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We have analyzed the expression and intracellular distribution, during oogenesis and embryogenesis, of Vg1 RBP, a protein implicated in the intracellular localization of Vg1 mRNA to the vegetal cortex of Xenopus oocytes. Vg1 RBP (protein) colocalizes with Vg1 RNA at all stages of oogenesis. Vg1 RBP RNA, however, localizes to the animal pole during late oogenesis, and remains in the animal blastomeres and ectodermal precursors until its zygotic transcription is activated, around stage 12. Vg1 RBP mRNA then becomes expressed throughout the neural epithelium. Vg1 RBP mRNA expression is also detected in what appears to be neural crest cells undergoing delamination and lateral migration. By tailbud stages, Vg1 RBP expression is present in the branchial arches, otic vesicle, pronephros, and along the neural tube. To examine the expression pattern in different species, we cloned the zebrafish homolog of Vg1 RBP by using a highly homologous EST clone to screen an embryonic cDNA library. In situ hybridization reveals that Vg1 RBP RNA localizes early in oogenesis to the animal pole. Although Vg1 RBP RNA is detected in all blastomeres of the early embryo, the expression pattern in the one day old zebrafish embryo is almost identical to that of the equivalent stage Xenopus embryo. These results indicate that the zygotic expression pattern is similar in frogs and fish, and that there is a conserved zygotic expression of Vg1 RBP distinct from its expression in the oocyte.

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Species referenced: Xenopus
Genes referenced: ddx6 gdf1 igf2bp3
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	Fig. 1. Vg1 RBP distribution in oocytes. (A) Vg1 RBP distribution in different stage oocytes was viewed by whole mount immunofluorescence staining with the anti B3 antibody and viewed by confocal microscopy. All of the oocytes are oriented with their vegetal poles down. Bar, 400 mm. (B) The distribution of Vg1 RNA (red channel) and Vg1 RBP (green channel) were compared in the same stage IV oocyte. Vegetal hemisphere, to the right.
	Fig. 2. Vg1 RBP protein and mRNA levels during development. (A) Western analysis of Xenopus proteins from two oocyte equivalents from each of the six oogenic stages. Vg1 RBP, anti-VgRBP antibody; Xp54, anti-Xp54 helicase antibody. (B) Four oocyte or embryo equivalents of total RNA from each of the indicated stages were probed for the presence of Vg1 RBP RNA. Stage 13 represents the approximate beginning of neurulation.
	Fig. 3. Vg1 RBP mRNA distribution in oocytes. Vg1 RBP mRNA is uniformly distributed throughout early stage oocytes, but becomes restricted to the animal hemisphere by stage V.
	Fig. 4. Vg1 RBP mRNA distribution in embryos. Whole mount in situ hybridization reveals the distribution of Vg1 RBP mRNA in embryos from stages 3 (4-cell stage, (A)); 20, (B); 26, (C); 30/31, (D,E). In (E), the embryo, following staining, was re®xed in Bouin's. In (B) the arrowheads indicate the area of putative neural crest delaminating and migrating from the midbrain (mb) and hindbrain. The cement gland (cg) is unlabeled. Ot, optic tectum; e, eye; ov, otic vesicle; ba, branchial arches; pn, pronephros; bi, blood islands.