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Cloning and Characterization of the Hemifacial Microsomia Related Gene.

半面畸形相关基因的克隆和表征。

基本信息

批准号：
10470456
负责人：
NAORA Hiroyuki
金额：
$ 7.1万
依托单位：
Shimane Prefectural Shimane Women's College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

To get an insight into the genetical predisposition and pathogenesis of the hemifacial microsomia (HFM), we analyzed the mutant locus of the HFM model mouse. By the P1 phage library screening, we isolated the chromosomal DNA from the mutant region. By the DNA sequencing, we screened the coding regions that corresponded to the mutant gene(s). At present, we sequenced about 10 kb, then we identified some regions that had highly homologous sequences with those of some known genes. These genes locate the different loci with HFM locus, then it is suggested that the gene(s) that responds to HFM pathogenesis is a nobel gene, and shares some domain structures to the known genes.We investigated the phenotype of homo-zygote of HFM mouse. By the natural mating between the hemi zygotes, we obtained 12 embryos at E9.5. Three embryos died as the rosy mass. PCR analysis showed that one of three embryos was a homo-zygote (another two embryos could not determine their genotypes due to DNA degradation). According to the size of the placenta, homo-zygotes embryo developed normally to E7.5. These results showed that the gene that disrupted in HFM mouse had an essential role for the normal development.The pathological screening was carried out at the late prenatal days (E13.5-17.5). In addition to the branchial arch anomalies that described previously, other anomalies and asymmetries at the maxilla, mandible, zygoma and facial soft tissue ware identified. These observation further supported that HFM mouse is a suitable animal model for the human HFM at the point of the pathological characteristics.

为了深入了解半侧颜面矮小症（HFM）的遗传易感性和发病机制，我们分析了HFM模型小鼠的突变位点。通过P1噬菌体文库筛选，我们从突变区中分离出染色体DNA。通过DNA测序，我们筛选出与突变基因对应的编码区。目前，我们已经测序了约10kb的基因序列，并发现了一些与已知基因高度同源的区域。这些基因与HFM基因位于不同的位点，表明与HFM发病机制相关的基因是一个nobel基因，并且与已知基因具有相同的结构域。通过半合子之间的自然交配，我们在E9.5获得了12个胚胎。三个胚胎死亡，呈玫瑰色团块。PCR分析显示，三个胚胎中有一个是纯合子（另外两个胚胎由于DNA降解而无法确定其基因型）。根据胎盘大小，纯合子胚胎发育正常至E7.5。这些结果表明，HFM小鼠中被破坏的基因对正常发育具有重要作用。除了先前所描述的鳃弓异常外，还发现了上颌骨、下颌骨、颌骨瘤和面部软组织的其他异常和不对称。这些观察结果进一步支持HFM小鼠是一种合适的人类HFM病理特征的动物模型。