title: Establishment of Gal4 transgenic zebrafish lines for analysis of development of cerebellar neural circuitry.
abstract: The cerebellum is involved in some forms of motor coordination and motor learning. Here we isolated transgenic (Tg) zebrafish lines that express a modified version of Gal4-VP16 (GFF) in the cerebellar neural circuits: granule, Purkinje, or eurydendroid cells, Bergmann glia, or the neurons in the inferior olive nuclei (IO) which send climbing fibers to Purkinje cells, with the transposon Tol2 system. By combining GFF lines with Tg lines carrying a reporter gene located downstream of Gal4 binding sequences (upstream activating sequence: UAS), we investigated the anatomy and developmental processes of the cerebellar neural circuitry. Combining an IO-specific Gal4 line with a UAS reporter line expressing the photoconvertible fluorescent protein Kaede demonstrated the contralateral projections of climbing fibers. Combining a granule cell-specific Gal4 line with a UAS reporter line expressing wheat germ agglutinin (WGA) confirmed direct and/or indirect connections of granule cells with Purkinje cells, eurydendroid cells, and IO neurons in zebrafish. Time-lapse analysis of a granule cell-specific Gal4 line revealed initial random movements and ventral migration of granule cell nuclei. Transgenesis of a reporter gene with another transposon Tol1 system visualized neuronal structure at a single cell resolution. Our findings indicate the usefulness of these zebrafish Gal4 Tg lines for studying the development and function of cerebellar neural circuits.
authors.affiliation: Laboratory of Organogenesis and Organ Function, Bioscience and Biotechnology Center, Nagoya University, Nagoya, Aichi 464-8601, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Takeuchi
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Aichi 464-8602, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Matsuda
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Aichi 464-8602, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Yamaguchi
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Asakawa
authors.affiliation: RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Miyasaka
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Lal
authors.affiliation: RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Yoshihara
authors.affiliation: Primate Research Institute, Kyoto University, Inuyama 464-8506, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Koga
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Kawakami
authors.affiliation: Laboratory of Organogenesis and Organ Function, Bioscience and Biotechnology Center, Nagoya University, Nagoya, Aichi 464-8601, Japan; Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Aichi 464-8602, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimizu
authors.affiliation: Laboratory of Organogenesis and Organ Function, Bioscience and Biotechnology Center, Nagoya University, Nagoya, Aichi 464-8601, Japan; Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Aichi 464-8602, Japan. Electronic address: hibi@bio.nagoya-u.ac.jp.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Hibi
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002465:Cell Movement; D002531:Cerebellum / Q000196:embryology* / Q000502:physiology; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009434:Neural Pathways; D009474:Neurons / Q000502:physiology; D011689:Purkinje Cells / Q000166:cytology; D013569:Synapses; D014157:Transcription Factors / Q000235:genetics*; D019076:Transgenes; D015027:Zebrafish / Q000196:embryology / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C542436:Gal4 protein, zebrafish; D014157:Transcription Factors; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins
keywords: Cell migration; Cerebellum; Gal4; Neural circuits; Zebrafish