权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Identification of the genes involved in the cell differentiation of the pancreas β cell and pre-clinical study of the diabetes model animal.

胰腺β细胞细胞分化相关基因的鉴定及糖尿病模型动物的临床前研究。

基本信息

批准号：
14370343
负责人：
KUSAKABE Moriaki
金额：
$ 8.9万
依托单位：
Institute for Animal Reproduction
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

The purpose of this experiment is to search the genes, which are involved in the hypertropic Island, and to examine the histological localization of the mRNA of these genes.As results, 1. We developed a novel high throughput in situ hybridization system to analyze the gene expression profiles steadily.2.The two-way subtraction libraries were made between the pancreas of 10 week-old KKAy and one of age matched BALB/cA. We selected 162 clones from KK-BALB library (K-B) and 153 from BALB-KK library (B-K) as specifically expressed clones. After homology analysis and clustering, 29 genes in K-B and 23 in B-K were identified. Next, quantitative RT-PCR analysis was done to select the clones. Finally, 11 KKAy specific including 1 unknown gene and 10 BALB specific genes including 2 unknown genes were selected. Reg I, Reg II and Reg III beta, which are involved in regeneration, were specifically enhanced their expression in the KKAy mouse. None of Reg genes was detected in 10 wks old BALB pancreas. All Reg genes were expressed only in acinar cells of 10 wks old KKAy pancreas, but not in the islet and the exocrine duct. However, in acinar cells of 27 wks old BALB pancreas, Reg I gene was strongly expressed. Reg II gene was faintly expressed in acinar cells. Reg III beta was hardly detectable. All Reg genes were strongly expressed only in acinar cells of 30 wks old KKAy. Of interest, the expression of Reg III beta was decreased in the cells surrounding the islet.3.Expression of KKAy specific genes were also examined in Streptozotocin induced type I diabetic mouse. As result, the expression of these genes was not specific in this model mouse.In conclusion, these data suggest that Reg genes may play important role in the beta cell regeneration in KKAy islet.

本实验的目的是寻找与高压岛相关的基因，并检测这些基因的组织学定位。结果：1.我们建立了一种新型的高通量原位杂交系统，以稳定地分析基因的表达谱。2.建立了10周龄KKAy和年龄匹配的BALB/CA胰腺的双向消减文库。我们从KK-BALB文库(K-B)中筛选出162个克隆，从BALB-KK文库(B-K)中筛选出153个克隆作为特异性表达克隆。经同源性分析和聚类分析，共鉴定出K-B基因29个，B-K基因23个。接下来，进行定量RT-PCR分析以筛选克隆。最终筛选出11个KKAy特异基因，包括1个未知基因；10个BALB特异基因，包括2个未知基因。参与再生的Reg I、Reg II和Reg III beta在KKAy小鼠中的表达被特异性地增强。10周龄BALB胰腺组织中未检测到REG基因。所有REG基因仅在10周龄KKAy胰腺的腺泡细胞中表达，而在胰岛和外分泌管中不表达。而在27周龄BALB胰腺的腺泡细胞中，Reg I基因呈强阳性表达。Reg II基因在腺泡细胞中有微弱表达。REG III测试版几乎检测不到。所有REG基因仅在30周龄KKAy的腺泡细胞中有强表达。3.链脲佐菌素诱导的I型糖尿病小鼠KKAy特异性基因的表达也被检测到。提示REG基因可能在KKAy胰岛的β细胞再生中起重要作用。