東邦大学 教育・研究業績データベース   
     


  イトウ マサノリ   Ito Masanori
  伊藤 雅方
   所属   東邦大学  医学部 医学科
   職種   講師
言語種別 英語
発表タイトル Fine analysis of regulatory region of the mouse Sry gene
会議名 29th International Conference on Animal Genetics
学会区分 国際学会
発表形式 ポスター掲示
講演区分 一般
発表者・共同発表者◎Ito M, Yokouchi K, Yoshida K, Kano K, Endo H, Hakamata Y, Miyazaki J, Naito K, Tojo H
発表年月日 2004/09/11
開催地
(都市, 国名)
Tokyo, Japan
概要 Sry (Sex determining region on Y chromosome) gene expression is restricted in undifferentiated gonads of mammals. There have been few studies on the regulatory elements of the Sry gene, mainly because no Sry-expressing cell lines have yet been established. We developed a useful tool for investigating the regulatory upstream region of Sry by means of the in vitro Cre/loxP system using primary cultured cells from CAG (cytomegalovirus enhancer and beta-actin promoter) /loxP/CAT/loxP/LacZ transgenic mice. We constructed a Sry/Cre fusion gene plasmid in which Cre expression is controlled by the 5’ and 3’ untranslated regions of mouse Sry that have been previously shown to induce testis development in XX transgenic mice. When Sry/Cre plasmids were transfected into the cells that had been prepared from the gonads, the brains and the livers of CAG/loxP/CAT/loxP/LacZ transgenic fetuses on 11.5 days post-coitus, only a small number of X-gal-stained cells were detected among the primary cultured cells from male and female gonads, while none were detected among the cells from the other tissues. The X-gal-positive cells were negative for alkaline phosphatase, indicating that these cells were somatic cells expressing Sry. Next, we constructed Sry/Cre plasmids with a differentially sized-upstream region of Sry. When Sry/Cre plasmid with 0.4kb upstream region of Sry was transfected into cells from various tissues, a large number of X-gal-positive cells were observed in the cells from tissues in which Sry is not expressed, indicating the loss of the stage- and tissue- specific expression of Sry. The Sry/Cre with a 0.5 kb upstream region maintained stage- and tissue-specific activity of Sry. The results suggested that the 0.4-0.5 kb upstream region of Sry is important for stage- and tissue- specific expression of Sry.