Analyses on the pathogenesis of hydrocephalus associated with L1CAM-mutation using in utero shRNA silencing

• Project/Area Number: 18390308
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Embryonic/Neonatal medicine
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: ITOH Kyoko Kyoto Prefectural University of Medicine, Department of Pathology and Applied Neurobiology, Associate Professor (80243301)
• Co-Investigator(Kenkyū-buntansha): FUSHIKI Shinji Kyoto Prefectural University of Medicine, Department of Pathology and Applied Neurobiology, Professor (80150572) ; IKEDA Hisafumi Osaka University, Department of Pharmaoology, lecturer (70322493)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥18,770,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥3,570,000); Fiscal Year 2007: ¥15,470,000 (Direct Cost: ¥11,900,000、Indirect Cost: ¥3,570,000); Fiscal Year 2006: ¥3,300,000 (Direct Cost: ¥3,300,000)
• Keywords: congenital malformation / L1CAM / Hydrocephalus / shRNA / in utero electroporation / L1cam / 大脳皮質形成 / PNA / レーザーマイクロダイセクション

Research Abstract

L1cam-silencing using in utero electroporation of shRNA for L1cam (L1-shRNA) was performed in order to gain insight into the pathogenesis of hydrocephalus associated with L1cam mutation. Transfection of L1-shRNA induced down-regulation of the expression of both L1cam mRNA and protein in cultured neuronal cells (Neuro2a) as compared to those transfected by negative control shRNA (shNC). When transfected to the cortical neurons that were cultured from mouse forebrain at embryonal 13.5 days (E13.5), L1-shRNA induced morphological changes, such as increased frequency of neurons with short and multiple neurites and abundant spines. L1-shRNA resulted in decreased length of total neurites and increased numbers of arborization as compared to those transfected by shNC. In utero electroporation of L1-shRNA was performed into the ventricular zone of murine forebrain at E13.5 days. On 2 to 4 days after transfection, the pattern of neuronal differentiation and migration showed no significant differences between L1-shRNA- and shNC-treated brains.

Research Products

• [Journal Article] Ionizing radiation downregulates ASPM, a gene responsible for microcephaly in humans (2008)
  • Author(s): Fujimori A, Yaoi T, Ogi H, Wang B, Suetomi K, Sekine E, Yu D, Kato T, Takahashi S, Okayasu R, Itoh K, Fushiki S.
  • Journal Title: Biochem Biophys Res Commun 369(3) Pages: 953-957
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed