Molecular genetics of SPINK5

SPINK5的分子遗传学

基本信息

批准号：
13470174
负责人：
YAMANISHI Kiyofumi
金额：
$ 10.37万
依托单位：
Hyogo College of Medicine (2003)Mie University (2002)Kyoto Prefectural University of Medicine (2001)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

Netherton syndrome is characterized by ichthyosis, atopy, and trichorrhexis invasinata. The disease gene for the syndrome has been identified to be SPINK5 for a serine protease inhibitor. However, the molecular pathogenesis of the ichthyosis is still unknown. This study is to analyze the role of serine protease systems in keratinization and elucidate the pathogenesis of ichthyosis using mouse models. The mouse SPINK5 gene was isolated from a 129Sv/J mouse genomic DNA library and made a targeting vector. After the homologous recombination in ES cells under the selection with G418, several ES clones with the disrupted SPINK5 gene were cloned. SPINK5 knockout mice were generated using one of those ES clones. However, no substantial abnormal phenotype was found in those knockout mice. On the other hand, transglutaminase 1 knockout mice(TGase1^<-/->) skin grafted onto nude mice developed an ichthyosiform skin phenotype. The gene expression profile of perinatal TGase1^<-/-> skin using a mouse DNA chip revealed that 38 genes are up-regulated and 236 genes are down-regulated in TGase1^<-/-> skin. Of those, BSSP, a gene for a serine protease, is expressed predominantly in brain and skin. Real-time PCR analysis showed that the BSSP gene is over-expressed in perinatal TGase1^<-/-> skin.Immunohistochemistry using an anti-BSSP antibody revealed that the fully developed icthyosiform skin of TGase1^<-/-> grafted skins are highly positive for BSSP. BSSP may be a target molecule for the formation of the ichthyosiform skin phenotype in keratinization disorders.

Netherton综合征的特征是鱼质病，atopy和Trichorrhexis Invasinata。综合征的疾病基因已被确定为丝氨酸蛋白酶抑制剂的Spink5。然而，鱼质病的分子发病机理仍然未知。这项研究是为了分析丝氨酸蛋白酶系统在角化化中的作用，并使用小鼠模型阐明了鱼质病的发病机理。从129SV/J小鼠基因组DNA文库中分离出小鼠Spink5基因，并制造了靶向载体。在使用G418选择的ES细胞中同源重组后，克隆了几个具有破坏Spink5基因的ES克隆。使用其中一种ES克隆生成Spink5敲除小鼠。然而，在那些敲除小鼠中未发现明显的异常表型。另一方面，跨谷氨酰胺酶1基因敲除小鼠（TGASE1^< - / - >）的皮肤接枝到裸鼠上的皮肤会形成鱼裂状的皮肤表型。使用小鼠DNA芯片皮肤的围产期TGASE1^< - / - >皮肤的基因表达谱显示38个基因被上调，并且在TGASE1^< - / - >皮肤中下调了236个基因。其中，BSSP是丝氨酸蛋白酶的基因，主要在大脑和皮肤中表达。实时PCR分析表明，使用抗BSSP抗体的围产期TGASE1^< - / - > SKIM.MMUNO组织化学中BSSP基因表达过表达，表明TGASE1^< - / - >移植的皮肤的完全发达的Icthyosiform皮肤对BSSP的阳性高度阳性。 BSSP可能是角质化疾病中鱼质皮肤表型形成的目标分子。