Elsevier

Gene Expression Patterns

Volume 5, Issue 2, December 2004, Pages 273-278
Gene Expression Patterns

Expression of the zebrafish Staufen gene in the embryo and adult

https://doi.org/10.1016/j.modgep.2004.07.007Get rights and content

Abstract

Staufen, a double stranded RNA binding protein, has been shown to be involved in creating and maintaining cellular asymmetry in the Drosophila oocyte, neuroblast, and mammalian neuron. Staufen binds to the 3′ UTR of specific mRNAs and acts in their localization and anchoring to various subcellular domains. Staufen's molecular interactions during development have been limited to investigations in Drosophila melanogaster. Since a vertebrate Staufen has not been studied in a developmental system, the aim of this study was to clone and characterize a staufen orthologue gene in the vertebrate developmental model, zebrafish. The zebrafish staufen-like sequence shows a 64% homology to the human staufen with a 81.2% homology in the highly conserved double stranded RNA binding domain (dsRBDs). Staufen maps on the LN54 radiation hybrid panel to linkage group 6, 16.25 cR from Z265 between fb22h06 and fi16e01.

Northern blot and in situ hybridization showed that staufen is expressed both maternally and zygotically. Zygotically expressed staufen is localized to the developing nervous system and at 24 h is highly concentrated in the subventricular zone of the developing brain. Maternally expressed staufen is dispersed in the mature oocyte and early embryo. In the adult, staufen is expressed in specific brain nuclei, the testis, neurons and Leydig cells.

Section snippets

Sequence analysis

A comparison of the zebrafish Staufen to other known sequences suggests it is an evolutionary intermediate between invertebrate and vertebrate sequences. The amino acid sequence of the Drosophila, human, and zebrafish Staufen are depicted in Fig. 1A and a representation of the double-stranded RNA binding domains (dsRBDs) and tubulin binding domains (TBD) in Fig. 1B The zebrafish staufen 5′ end includes the first dsRBD, which has a 64% identity to Drosophila and is missing in human staufen;

In situ hybridization (whole mount and sectioned tissue)

Whole mount in situ hybridization was performed as previously described (Thisse et al., 1994). In situ hybridization was performed on zebrafish sectioned tissue as previously described (Braat et al., 1999). The anti-sense staufen probe was a 1600 pb fragment produced by linearizing the TOPO TA vector (Invitrogen) with Bam HI and using T7 RNA polymerase for in vitro transcription. The sense probe was linearized with Hind III and transcribed using T3 polymerase. Vasa in situ hybridization on early

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  • Cytoplasmic remodelling and the acquisition of developmental competence in pig oocytes

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    Citation Excerpt :

    Among the many RNA-binding proteins identified so far, Staufen has been shown to be involved in RNA localization in Drosophila oocytes (Gavis and Lehmann, 1992; Hachet and Ephrussi, 2004) and Xenopus laevis eggs (Yoon and Mowry, 2004). Expression of Staufen has also been recently demonstrated in the zebrafish embryo (Bateman et al., 2004). The pig orthologue for Staufen has been recently identified (Staufen; EMBL accession number: AJ969068) in our laboratory (Brevini et al., 2005b).

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