Search for genes involved in neuronal migration in cerebral cortex of X-irradiated embryonic mice

点击翻译按钮获取中文摘要

On the basis of our previous studies that prenatal low-level ionizing radiation impedes neuronal migration in rodent cerebral cortices, we have attempted to search for genes involved in neuronal migration by using RLCS (Restriction Landmark cDNA Scanning) method.The samples for RLCS were obtained from the whole brains of embryonic day 14.5 mice ; one group X-irradiated at 0.5 Gy, the other non-irradiated, We have so far made screening on total 12,000 spots by employing 10 restriction enzymes. After comparing the RLCS profiles from irradiated brains with those from non-irradiated brains we found that among the spots obtained 0.07% showed up-regulation and 0.11% down-regulation. From the spots showing down-regulation we succeeded in sequencing one of the down-regulated spots. The gene codes for one of the cell adhesion molecule like protein comprising of 295 to 306 amino acids, which belongs to the immunoglobulin superfamily, although the function of the molecule remains unknown. By applying real-time PCR method, we have confirmed that the expression level of the gene in X-irradiated brains was suppressed down to 30-40% of the non-irradiated control.We have also attempted to devise a new method, called "single cell RLCS" that enables us to study the changes of gene expression at single cell level. During the term of the present research project we have completed in establishing single cell RLCS from the technical viewpoint.