Ben-Dror I, Havazelet N, Vardimon L
Department of Biochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1117-21. doi: 10.1073/pnas.90.3.1117.
In chicken embryo retina, competence for induction of the glutamine synthetase [L-glutamate:ammonia ligase (ADP-forming); EC 6.3.1.2] gene by glucocorticoid hormones increases progressively with development; this competence is minimal in 6-day retina (E6) and high by day 10 (E10). Because the level of glucocorticoid receptors (GRs) in the retina does not increase during that time, we investigated whether the transcriptional activity of GR increased between days 6 and 10 of development. The glucocorticoid-inducible chloramphenicol acetyltransferase (CAT) constructs 2GRE-37TK and p delta G46TCO, which contain glucocorticoid-responsive elements attached to a TATA box and to the thymidine kinase promoter, respectively, were transfected into E6 and E10 retinas, and their inducibility was examined. CAT expression could be induced in the transfected E10 retina but was not induced in the transfected E6 retina. However, induction was obtained also in E6 retina after cotransfection with a GR expression vector. Noninducible CAT constructs (pRSV-CAT, pSV2CAT, and pBLCAT2) were expressed at both ages at similar levels. The CAT construct pGS2.1CAT, which is controlled by the upstream sequence of the chicken glutamine synthetase gene, could be induced in E10 retina but was not induced in E6 retina; however, cotransfection with the GR expression vector resulted in induction of pGS2.1CAT also in E6 retina. We interpret these results as showing that the transcriptional activity of GR in embryonic retina is developmentally controlled and suggest that its increase is causally implicated in the development of competence for glutamine synthetase induction.
在鸡胚视网膜中,糖皮质激素诱导谷氨酰胺合成酶[L-谷氨酸:氨连接酶(ADP形成);EC 6.3.1.2]基因的能力随发育进程逐渐增强;这种能力在6日龄视网膜(E6)中最低,到10日龄(E10)时则很高。由于在此期间视网膜中糖皮质激素受体(GRs)的水平并未增加,我们研究了GR的转录活性在发育的第6天至第10天之间是否有所增加。分别含有与TATA盒和胸苷激酶启动子相连的糖皮质激素反应元件的糖皮质激素诱导型氯霉素乙酰转移酶(CAT)构建体2GRE-37TK和pδG46TCO被转染到E6和E10视网膜中,并检测它们的诱导性。CAT表达可在转染的E10视网膜中被诱导,但在转染的E6视网膜中未被诱导。然而,与GR表达载体共转染后,E6视网膜中也获得了诱导。不可诱导的CAT构建体(pRSV-CAT、pSV2CAT和pBLCAT2)在两个年龄段的表达水平相似。由鸡谷氨酰胺合成酶基因上游序列控制的CAT构建体pGS2.1CAT在E10视网膜中可被诱导,但在E6视网膜中未被诱导;然而,与GR表达载体共转染导致pGS2.1CAT在E6视网膜中也被诱导。我们将这些结果解释为表明胚胎视网膜中GR的转录活性受到发育调控,并表明其增加与谷氨酰胺合成酶诱导能力的发育存在因果关系。