Elsevier

Gene Expression Patterns

Volume 6, Issue 8, 2 October 2006, Pages 761-771
Gene Expression Patterns

Cloning and embryonic expression of five distinct sfrp genes in the zebrafish Danio rerio

https://doi.org/10.1016/j.modgep.2006.01.006Get rights and content

Abstract

Recently, a new member of the secreted frizzled-related protein (sFRP) family, named tlc, has been identified as expressed by the anterior neural border (ANB) cells in the zebrafish Danio rerio. Tlc plays an important role in telencephalic induction and patterning. In absence of Tlc, formation of the telencephalon is severely delayed, but not abolished. This prompted us to clone the other zebrafish sfrp family members and analyse their expression patterns, in search of a family member that may partly functionally overlap with Tlc. Except sizzled, expression profile of sfrp genes in zebrafish has not been reported so far. Here, we describe the cloning of full-length cDNA for sfrp1a, sfrp1b, sfrp2, sfrp3 and sfrp5 gene transcripts and we examine their expression at different embryonic stages. Only sfrp1a is expressed in the anterior neural plate including the ANB cells where and when tlc is expressed. Interestingly, compared to both tlc and sfrp1a, wnt genes are complementary expressed more posteriorly in the neural plate. Later, both sfrp1a and sfrp5 expression profiles are overlapping, in particular at pharyngula stage these genes are expressed in the ventral part of the forebrain, midbrain and hindbrain. sfrp1b, sfrp2 and sfrp3 are mainly expressed in mesodermal and endodermal embryonic tissues. Expression profiles of these different genes in zebrafish gave interesting clues on the possible function and evolution of sFRPs in zebrafish and other organisms.

Section snippets

Phylogenic distribution of sFRPs

To date, eight distinct sFRPs have been identified, mostly in vertebrates (i.e., amphibians, birds, fishes and mammals), but also in the urochordate Ciona intestinalis (Dehal et al., 2002). One sFRP has been characterized in the sea urchin Strongylocentrotus purpuratus (Illies et al., 2002). Finally, no sFRP has been identified so far in protostome organisms, suggesting that sFRP arose after the separation between protostome and deuterostome lineages. In silico analyses of amino-acid sequences

Cloning of sfrp genes

Except for the gene sfrp3 (gift from M. Furutani-Seiki), the full-length cDNA sequence of sfrp1a, sfrp1b, sfrp2 and sfrp5 was obtained by PCR amplification using a cDNA library constructed using the Invitrogen cDNA single strand RT-PCR kit. Total RNA is extracted from 24 hpf zebrafish embryo using the Trizol reagent (Invitrogen). For the PCR amplification, oligonucleotides are synthesized using information obtained on sfrp gene sequences from the zebrafish genome database Ensembl available on

Acknowledgements

We are thankful to Dr. Jamile Hazan for the cloning of the sfrp1b gene. This study was supported by grants from the Wellcome trust and the Medical Research Council awarded to C.H.

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