RESUMEN
Objective: To explore the molecular evolution of zebrafish rnf141 gene and its expression in the embryo and adult zebrafish. Methods: The molecular characteristics of zebrafish rnf141 gene were analyzed through a bioinformatics approach. Expression of rnf141 gene in the embryos of different developmental stages and tissues of adult fish were examined by whole-mount in situ hybridization and multiple-tissue RT-PCR technique. Results: We noticed that zebrafish RNF141 protein shared a 79.8% and 78.5% homologies to its homologues in human and mouse (ZNF230, Znf230), respectively, especially that the C3HC4-type domain and the same conserved cysteine and histidine residues shared a 84.8% and 100% homologies to its homologues in human or mouse. Moreover, the secondary structure of C3HC4-type domain in zebrafish RNF141 protein was the same as that in its human homologue. Whole-mount in situ hybridization showed that zebrafish rnf141 gene expressed ubiquitously during embryogenesis, with higher expression found in the telencephalon, cerebellum and hindbrain. RT-PCR revealed rnf141 gene expression in all the adult tissues detected, and abundance was observed in the testicular tissue. Conclusion: These data suggest that zebrafish rnf141 gene is conservative in the vertebrate and might play important roles in embryogenesis and spermatogenesis.
RESUMEN
ZNF230 is a novel zinc finger gene cloned by our laboratory. In order to understand the potential functions of this gene in vertebrate development, we cloned the zebrafish orthologue of human ZNF230, named rnf141. The cDNA fragment of rnf141 was obtained by rapid amplification of cDNA ends (RACE). The open reading frame (ORF) encodes a polypeptide of 222 amino acids which shares 75.65 percent identity with the human ZNF230. RT-PCR analysis in zebrafish embryo and adult tissues revealed that rnf141 transcripts are maternally derived and that rnf141 mRNA has a broad distribution. Zygotic rnf141 message is strongly localized in the central nervous system, as shown by whole-mount in situ hybridization. Knockdown and over expression of rnf141 can induce abnormal phenotypes, including abnormal development of brain, as well as yolk sac and axis extendsion. Marker gene analysis showed that rnf141 may play a role in normal dorsoventral patterning of zebrafish embryos, suggesting that rnf141 may have a broad function during early development of vertebrates.