title: Identification and characterization of the RNA-binding protein Rbfox3 in zebrafish embryo.
abstract: Rbfox3, an RNA-binding fox protein, binds to the antibody to pan-neuronal marker, neuronal nuclei (NeuN). Rbfox3 is expressed in neural tissues across a wide range of species including mammals, birds, and amphibians. However, the molecular identity of Rbfox3 in the zebrafish is largely unknown. In this study, we cloned two zebrafish Rbfox3 genes, Rbfox3a and Rbfox3b. We also cloned the Rbfox3-d31 isoform, which excludes a 93-nucleotide alternative exon within the RNA-recognition motif in both, Rbfox3a and Rbfox3b. Multiple protein sequence alignment revealed that the amino acid sequence for residues 1-20 of the zebrafish Rbfox3, which is the epitope region of NeuN antibody, was different from that of other species. Therefore, NeuN antibody lost its function as a neuronal marker antibody in zebrafish. Reverse transcriptase-polymerase chain reaction showed that both Rbfox3-d31 transcripts were abundant in the early blastula stage, after which they dramatically reduced, suggesting that these isoforms exist mainly as maternal transcripts. In contrast, full-length Rbfox3 transcripts were detected from the 24 h post-fertilization embryo, expression was also maintained at a constant level. Furthermore, full-length Rbfox3-expressing cells were located within the central nervous system during later stages of the zebrafish embryo. Our study provides insight into the molecular structure of zebrafish Rbfox3 as a step towards genetic association studies investigating the developmental role of Rbfox3.
authors.affiliation: Department of Pharmacology, College of Medicine, Chungnam National University, Daejeon, 35015, Republic of Korea.
authors.forename: Minho
authors.initials: M
authors.lastname: Won
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, 34134, Republic of Korea.
authors.forename: Siyeo
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, 34134, Republic of Korea.
authors.forename: Sunkyung
authors.initials: S
authors.lastname: Choi
authors.affiliation: Department of Biological Sciences, College of Bioscience and Biotechnology, Chungnam National University, Daejeon, 34134, Republic of Korea.
authors.forename: Hyunju
authors.initials: H
authors.lastname: Ro
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, 34134, Republic of Korea.
authors.forename: Ki-Jung
authors.initials: KJ
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, School of Medicine, Institute of Health Sciences, Gyeongsang National University, Jinju, 52727, Republic of Korea.
authors.forename: Jung-Hwan
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, 34134, Republic of Korea.
authors.forename: Kyoon Eon
authors.initials: KE
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, 34134, Republic of Korea. Electronic address: kimkk@cnu.ac.kr.
authors.forename: Kee K
authors.initials: KK
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D008969:Molecular Sequence Data; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D016601:RNA-Binding Proteins; D029961:Zebrafish Proteins; C000609590:rbfox3a protein, zebrafish
keywords: Alternative splicing; NeuN; RNA-binding protein; Zebrafish Rbfox3