Abdallah B, Sachs L, Hourdry J, Wegnez M, Denis H, Demeneix B, Mazabraud A
Centre de Génétique Moléculaire, Université Pierre et M. Curie (Paris VI), Gif-sur-Yvette, France.
Int J Dev Biol. 1996 Apr;40(2):507-14.
In situ hybridization was used to follow the distribution of the mRNAs encoding the somatic form of elongation factor 1 alpha (EF-1 alpha S) and the germinal counterparts of this factor, thesaurin a and EF-1 alpha O, throughout metamorphosis in the gonads of Xenopus laevis tadpoles. EF-1 alpha S mRNA is detected before metamorphosis in both the somatic and germ cells of the gonads. In contrast, thesaurin a and EF-1 alpha O mRNAs are first detected in spermatogonia and oogonia at stages 60-62, corresponding to the climax of metamorphosis and to the peak of circulating thyroid hormone. To determine whether thyroid hormone, the instigator of metamorphosis, is involved in regulating the expression of the germinal gene EF-1 alpha O, Xenopus XTC cells were transfected with an EF-1 alpha O promoter sequence inserted in front of the luciferase reporter gene. Addition of T3 to the cell culture medium induced a dose-dependent increase in transcription from the EF-1 alpha O promoter. This effect was enhanced when the construct was cotransfected with an expression vector for a Xenopus thyroid hormone receptor. Our data show that germ cells switch from a somatic to a germ-cell specific mode of expression during metamorphosis. Furthermore, this switch appears to be induced by thyroid hormone.
采用原位杂交技术追踪非洲爪蟾蝌蚪性腺在整个变态发育过程中,编码延伸因子1α体细胞形式(EF-1αS)及其生殖细胞对应物thesaurus a和EF-1αO的mRNA的分布情况。在变态发育前,性腺的体细胞和生殖细胞中均可检测到EF-1αS mRNA。相比之下,thesaurin a和EF-1αO mRNA首次在第60 - 62阶段的精原细胞和卵原细胞中被检测到,这一阶段对应变态发育的高峰期以及循环甲状腺激素的峰值。为了确定变态发育的引发剂甲状腺激素是否参与调节生殖细胞基因EF-1αO的表达,将插入荧光素酶报告基因前的EF-1αO启动子序列转染至非洲爪蟾XTC细胞中。向细胞培养基中添加T3可诱导EF-1αO启动子转录呈剂量依赖性增加。当该构建体与非洲爪蟾甲状腺激素受体的表达载体共转染时,这种效应会增强。我们的数据表明,生殖细胞在变态发育过程中从体细胞特异性表达模式转变为生殖细胞特异性表达模式。此外, 这种转变似乎是由甲状腺激素诱导的。