期刊
FASEB JOURNAL
卷 26, 期 11, 页码 4662-4674出版社
FEDERATION AMER SOC EXP BIOL
DOI: 10.1096/fj.12-207530
关键词
PC12; synaptic vesicles
资金
- Ministry of Education, Culture, Sports, Science, and Technology, Japan [21390050, 21590210]
- Kato Memorial Bioscience Foundation
- Mishima Kaiun Memorial Foundation
- Grants-in-Aid for Scientific Research [24390046, 21590210, 21390050, 24659084, 23591347] Funding Source: KAKEN
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.-Hashimoto, Y., Muramatsu, K., Kunii, M., Yoshimura, S., Yamada, M., Sato, T., Ishida, Y., Harada, R., Harada, A. Uncovering genes required for neuronal morphology by morphology-based gene trap screening with a revertible retrovirus vector. FASEB J. 26, 4662-4674 (2012). www.fasebj.org
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