Abnormal gene expressions in the Gli3 and Shh of the mouse homolog of GCPS, Pdn

GCPS、Pdn 小鼠同系物的 Gli3 和 Shh 基因表达异常

• 批准号: 12670015
• 负责人: NARUSE Ichiro
• 金额: $ 2.5万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670015/
• 关键词: Gli3; Shh; Pdn; GCPS; Pdnマウス; 多指症; 無嗅脳症

Phenotype of genetic polydactly/archinencephaly mouse (Pdn/Pdn) is similar to Greig cephalopolysyndactyly syndrome (GCPS), whose responsible gene is GLI3. Suppression of Gli3 gene expression had been observed in Pdn/Pdn. So, the responsible gene of Pdn/Pdn has been considered to be Gli3, but mutation point has not been demarcated.In this research project, it was determined that 5442 of a transposon was inserted in the position 41286/41287 of intron 3 of Gli3 gene in Pdn mouse. This transposon has 317-bp long terminal repeats in both ends. Forward and reverse PCR primers were constructed in the intron 3 near the insertion point, and forward primer in the transposon was also a constructed. Then we could discriminate +/+, Pdn/+, Pdn/Pdn embryos with the PCR product by these primers. Using this rapid PCR method, we can determine the genotypes of the young embryos.Gli3 gene expression in Pdn/+ was suppressed to about 60% for +/+, and that in Pdn/Pdn was 20-30% for +/+ through all the embryonic and neonatal periods examined, but stage dependency of gene expression was not observed. Shh has been considered to be antagonist of Gli3. So, we speculated that Shh would be overexpressed in the Pdn/Pdn mouse embryos whose Gli3 gene expression was suppressed. Shh expression was different among genotypes only by on day of 9 of gestation.

遗传性多指/无脑畸形小鼠（Pdn/Pdn）的表型与Greig头多并指综合征（GCPS）相似，其致病基因为GLI 3。在Pdn/Pdn中观察到Gli 3基因表达的抑制。本研究确定Pdn小鼠Gli 3基因内含子3的41286/41287位置插入了5442个转座子。该转座子在两端具有317-bp长的末端重复序列。在第3内含子插入点附近构建正向和反向PCR引物，并在转座子中构建正向引物。利用这些引物的PCR产物可以区分+/+、Pdn/+、Pdn/Pdn胚胎。结果表明，Gli 3基因在Pdn/+和Pdn/Pdn中的表达量分别为60%和20-30%，但未观察到基因表达的阶段依赖性。Shh被认为是Gli 3的拮抗剂。因此，我们推测Shh将在Gli 3基因表达被抑制的Pdn/Pdn小鼠胚胎中过表达。Shh基因型之间的表达差异仅在妊娠第9天。

项目成果

Naruse I., Ueta E.: "Hydrocephalus manifestation in the genetic arhinencephaly mouse (Pdn)"Cong.Anom.. 42:1. 27-31 (2002)

Naruse I.、Ueta E.：“遗传性无脑畸形小鼠 (Pdn) 中的脑积水表现”Cong.Anom.. 42:1。

成瀬 一郎: "形の科学辞典"形の科学会編, 朝倉書店(未定). (2003)

成濑一郎：《形式科学词典》，形式科学学会编，朝仓书店（待定）（2003 年）。

Naruse I.,Keiro H.,Yamada Y.and Masaki S.: "Mechanism of polydactyly manifestation in mice and its extrapolation to humans."Cong.Anom.. 40. 300-308 (2000)

Naruse I.、Keiro H.、Yamada Y. 和 Masaki S.：“小鼠多指畸形表现的机制及其对人类的推断。”Cong.Anom.. 40. 300-308 (2000)

Tanaka T., Naruse I., Hongo T., Xu M-J., Nakahata T. and Ishii S.: "Extensive brain hemorrhage and embryonic lethality in a mouse null mutant of GREB-binding protein"Mech. Develop.. 95. 133-145 (2000)

Tanaka T.、Naruse I.、Hongo T.、Xu M-J.、Nakahata T. 和 Ishii S.：“GREB 结合蛋白的小鼠无效突变体导致广泛脑出血和胚胎致死”Mech。

Naruse I., Keino H., Yamada Y., Masaki S., Hui C-C.: "Phenotypic differences in the brains and limb of mutant mice caused by differences of Gli3 gene expression levels"Cong. Anom.. 41:2. 89-94 (2001)

Naruse I.、Keino H.、Yamada Y.、Masaki S.、Hui C-C.：“Gli3 基因表达水平差异导致突变小鼠大脑和肢体的表型差异”