Abstract
Robo2, a member of the robo gene family, functions as a repulsive axon guidance receptor as well as a regulator of cell migration and tissue morphogenesis in different taxa. In this study, a novel isoform of the zebrafish robo2 (robo2_tv2), which included an otherwise alternatively spliced exon (CAE), has been characterized. Robo2_tv2 is expressed differentially in most non-neuronal tissues of adult zebrafish whereas robo2_tv1 expression to a great extent is restricted to the brain and eye. In zebrafish, robo2_tv2 exhibits a very-low-level basal expression starting from 1 day post fertilization until the mid-larval stages, at which time its expression increases dramatically and could be detected throughout adulthood. Our findings demonstrate that the amino acid sequence coded by CAE of the robo2 gene is highly conserved between zebrafish and mammals, and also contains conserved motifs shared with robo1 and robo4 but not with robo3. Furthermore, we provide an account of differential transcription of the CAE homolog in various tissues of the adult rat. These results suggest that the alternatively spliced robo2 isoforms may exhibit tissue specificity.
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Acknowledgements
The authors thank Dr. Kristen Ward and Rasit Ozturk for helpful comments. This research was supported by grants from The Scientific and Technical Research Council of Turkey, TUBITAK (TBAG-2285, 103T03) and Bilkent University to O. Konu. Ertugrul Dalkic has a graduate scholarship from TUBITAK.
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Communicated by M. Hammerschmidt
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Supplement 1
a The exon/intron structure of ENSDART00000014877 which corresponds to 3’ of zebrafish robo2 is shown in detail (www.ensembl.org). The putative exons (PE) present in the Genscan but not found in the reported transcript are highlighted with open boxes; the conserved alternative exon (CAE) is highlighted in a bold open box b The exon/intron structure of Ensembl transcripts corresponding to 3’ of rat robo2 is shown in detail (www.ensembl.org). The conserved alternative exon (CAE) is highlighted in a bold open box
Supplement 2
Robo2 primer pair conditions used for RT-PCR (PDF 39 kb)
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Dalkic, E., Kuscu, C., Sucularli, C. et al. Alternatively spliced Robo2 isoforms in zebrafish and rat. Dev Genes Evol 216, 555–563 (2006). https://doi.org/10.1007/s00427-006-0070-y
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DOI: https://doi.org/10.1007/s00427-006-0070-y