title: Uncovering genes required for neuronal morphology by morphology-based gene trap screening with a revertible retrovirus vector.
abstract: The molecular mechanisms of neuronal morphology and synaptic vesicle transport have been largely elusive, and only a few of the molecules involved in these processes have been identified. Here, we developed a novel morphology-based gene trap method, which is theoretically applicable to all cell lines, to easily and rapidly identify the responsible genes. Using this method, we selected several gene-trapped clones of rat pheochromocytoma PC12 cells, which displayed abnormal morphology and distribution of synaptic vesicle-like microvesicles (SLMVs). We identified several genes responsible for the phenotypes and analyzed three genes in more detail. The first gene was BTB/POZ domain-containing protein 9 (Btbd9), which is associated with restless legs syndrome. The second gene was cytokine receptor-like factor 3 (Crlf3), whose involvement in the nervous system remains unknown. The third gene was single-stranded DNA-binding protein 3 (Ssbp3), a gene known to regulate head morphogenesis. These results suggest that Btbd9, Crlf3, and Ssbp3 regulate neuronal morphology and the biogenesis/transport of synaptic vesicles. Because our novel morphology-based gene trap method is generally applicable, this method is promising for uncovering novel genes involved in the function of interest in any cell lines.
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Muramatsu
authors.forename: Masataka
authors.initials: M
authors.lastname: Kunii
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Yoshimura
authors.forename: Minami
authors.initials: M
authors.lastname: Yamada
authors.forename: Takashi
authors.initials: T
authors.lastname: Sato
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Ishida
authors.forename: Reiko
authors.initials: R
authors.lastname: Harada
authors.forename: Akihiro
authors.initials: A
authors.lastname: Harada
mesh_terms: D000818:Animals; D001427:Bacterial Toxins; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005786:Gene Expression Regulation / Q000502:physiology*; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors; D059785:Karyotype; D016254:Mutagenesis, Insertional / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D016716:PC12 Cells; D052899:Pore Forming Cytotoxic Proteins; D034741:RNA, Small Interfering; D051381:Rats; D012190:Retroviridae; D014157:Transcription Factors
chemical_list: D001427:Bacterial Toxins; D052899:Pore Forming Cytotoxic Proteins; D034741:RNA, Small Interfering; D014157:Transcription Factors; C032676:aerolysin