Study of the gene expression in development and regulation of methylation of DNA

DNA甲基化发育及调控中基因表达的研究

• 批准号: 01540590
• 负责人: YASUGI Sadao
• 金额: $ 1.28万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01540590/
• 关键词: regulation of gene expression; pepsinogen gene; embryonic development; methylation of DNA

The aim of the present study is to elucidate the regulation mechanisms of genes which show stage-specific and organ-specific expression during animal development. I used for this purpose the embryonic chicken pepsinogen (ECPg) gene which is expressed specifically in the epithelial cells of proventriculus (glandular stomach) during chicken embryonic period.In mouse and human, degree of methylation in DNA of pepsinogen genes has shown to become lower in the organs expressing pepsinogens. So I examined the degree of methylation in ECPg gene obtained from ECPg-expressing and ECPg-non-expressing organs.ECPg gene showed rather wide range polymorphisms when genomic DNA from chicken organs was digested with several restriction enzymes and subjected to Southern hybridization with cDNA of ECPg as a probe. I selected, therefore, the individuals which had the same hybridization pattern for further analysis. DNAs from proventriculus, gizzard, small intestine and muscle showed the same hybridization pattern when they were digested with methylation-sensitive restriction endonucleases such as Hha I or Hpa II. These results indicate that, ECPg expresssion is not regulated by methylation of DNA itself, but by sis- or trans-acting factors in the proventricular epithelial cells.

本研究的目的是阐明在动物发育过程中具有阶段特异性和器官特异性表达的基因的调控机制。为此，我使用了鸡胚胃蛋白酶原（ECPg）基因，该基因在鸡胚胎期腺胃上皮细胞中特异性表达，在小鼠和人中，胃蛋白酶原基因DNA的甲基化程度在表达胃蛋白酶原的器官中显示出降低。因此，本研究检测了ECPg表达器官和非表达器官中ECPg基因的甲基化程度，发现ECPg基因在鸡各器官基因组DNA中表现出较广泛的多态性，并以ECPg cDNA为探针进行Southern杂交。因此，我选择了具有相同杂交模式的个体进行进一步分析。用甲基化敏感的限制性内切酶Hha Ⅰ或Hpa Ⅱ消化前胃、肌胃、小肠和肌肉的DNA，结果显示出相同的杂交模式。这些结果表明，ECPg的表达不受DNA本身甲基化的调节，而是受腺室上皮细胞中顺式或反式作用因子的调节。

项目成果

YASUGI, S.: "Pepsinogenーlike immunoreactivity in ascidian stomach and intestine: Immunohistochemical and biochemical study." Zool. Sci.6. 283-288 (1989)

YASUGI, S.：“海鞘胃和肠中的胃蛋白酶原样免疫反应性：免疫组织化学和生化研究。Zool。”283-288（1989）。

YASUGI,S.: "Mesenchymalーepithelial interactions in the organogenesis of digestive tract." Zool.Sci.7. 159-170 (1990)

YASUGI, S.：“消化道器官发生中的间质-上皮相互作用。”Zool.Sci.7（1990）。

YASUGI,S.: "Mesenchymal-epithelial interactions in the organogenesis of digestive tract." Zool.Sci.6. (1990)

YASUGI,S.：“消化道器官发生中的间质-上皮相互作用。”

YASUGI, S.: "Mesenchymalーepithelial interactions in the organogenesis of digestive tract." Zool. Sci.7. 159-170 (1990)

YASUGI, S.：“消化道器官发生中的间充质-上皮相互作用。”Sci.7 159-170（1990）。

TAKIGUCHIーHAYASHI, K.: "Transfilter analysis of the inductive influence of proventricular mesenchyme on stomach epithelial differentiation of chick embryos." Roux' ArcH. Develop. Biol.198. 460-466 (1990)

TAKIGUCHI-HAYASHI, K.：“前室间质对鸡胚胎胃上皮分化的诱导影响的分析。”Roux Biol.198（1990）。