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

点击翻译按钮获取中文摘要

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.