Patejunas G, Young A P
Mol Cell Biol. 1987 Mar;7(3):1070-7. doi: 10.1128/mcb.7.3.1070-1077.1987.
We have isolated a glutamine synthetase cDNA clone derived from chicken retinal RNA. The clone detects a 3.2-kilobase RNA in chicken retina, liver, and brain, based on Northern blotting analysis. The dramatic developmental rise observed for the retinal enzyme, assayed as glutamyl transferase activity, is accompanied by a corresponding rise in this RNA. Injection of hydrocortisone 21-phosphate into the yolk sac of day 10 embryos produces an increase in retinal glutamine synthetase mRNA and glutamyl transferase activity, assayed 4 days after injection. An increase in glutamine synthetase mRNA is also observed within 2 h of incubation of retinal organ cultures with hydrocortisone. Moreover, incubation of these cultures with cycloheximide at a concentration that inhibits protein synthesis by 93% affects neither the basal level nor the hydrocortisone-mediated induction of glutamine synthetase mRNA. Although expression of this RNA is developmentally regulated in the brain, steroid hormone injection does not result in a substantial induction. Hepatic glutamine synthetase mRNA is expressed constitutively between embryonic day 10 and 6 days after hatching and is also not hormone inducible. Southern blotting data with chicken DNA digested with EcoRI, HindIII, and BamHI are best interpreted in terms of the cDNA clone detecting only one gene. If so, several cell-type-specific regulatory mechanisms must function to modulate expression of this gene during development.
我们从鸡视网膜RNA中分离出了一个谷氨酰胺合成酶cDNA克隆。根据Northern印迹分析,该克隆在鸡视网膜、肝脏和大脑中检测到一种3.2千碱基的RNA。以谷氨酰转移酶活性测定的视网膜酶在发育过程中显著升高,与此同时这种RNA也相应增加。在第10天胚胎的卵黄囊中注射21-磷酸氢化可的松,在注射后4天测定,视网膜谷氨酰胺合成酶mRNA和谷氨酰转移酶活性增加。在用氢化可的松培养视网膜器官培养物的2小时内,也观察到谷氨酰胺合成酶mRNA增加。此外,用环己酰亚胺以抑制蛋白质合成93%的浓度培养这些培养物,既不影响基础水平,也不影响氢化可的松介导的谷氨酰胺合成酶mRNA的诱导。虽然这种RNA的表达在大脑中受到发育调控,但注射类固醇激素并不会导致大量诱导。肝谷氨酰胺合成酶mRNA在胚胎第10天到孵化后6天之间持续表达,也不受激素诱导。用EcoRI、HindIII和BamHI消化鸡DNA的Southern印迹数据,最好根据仅检测到一个基因的cDNA克隆来解释。如果是这样,在发育过程中必须有几种细胞类型特异性调节机制发挥作用来调节该基因的表达。