Piperberg J B, Reif-Lehrer L
Cell Biophys. 1984 Jun;6(2):131-48. doi: 10.1007/BF02788592.
Glutamine synthetase (GS) activity is enhanced in cultured whole retinas when a 72 h incubation at 37 degrees C is preceded by storage at 4 degrees C for 2-24 h. This enhancement occurs even in the absence of glucocorticoids and is maximal in retinas from 11 to 14 d embryos. In comparison, cortisol-induced increases in retinal GS activity at 37 degrees C are optimal in retinas from 8 to 12 d embryos. This study, using cycloheximide (an inhibitor of protein synthesis) and cordycepin (an inhibitor of RNA synthesis), indicates that both protein and RNA synthesis are required for the 4 degrees C storage enhancement of GS activity. The necessary RNA synthesis occurs within the first 48 h following transfer to 37 degrees C and does not require concomitant protein synthesis. Uridine uptake, but not incorporation into trichloroacetic acid-precipitable material, is increased by initial 4 degrees C storage when compared with whole retina controls incubated at 37 degrees C for the total time. In contrast, both uptake and incorporation of amino acids are increased in 4 degrees C-stored retinas for as long as 72 h subsequent to transfer from 4 to 37 degrees C. This suggests that enhancement GS activity may arise from a combination of elevated general protein synthesis and specific messenger-RNA synthesis following 4 degrees C storage.
当在37℃孵育72小时之前先在4℃储存2 - 24小时时,培养的整个视网膜中的谷氨酰胺合成酶(GS)活性会增强。即使在没有糖皮质激素的情况下这种增强也会发生,并且在11至14天胚胎的视网膜中增强最为明显。相比之下,在37℃时,皮质醇诱导的视网膜GS活性增加在8至12天胚胎的视网膜中最为显著。本研究使用环己酰亚胺(一种蛋白质合成抑制剂)和虫草素(一种RNA合成抑制剂)表明,蛋白质和RNA合成对于4℃储存增强GS活性都是必需的。必要的RNA合成发生在转移到37℃后的前48小时内,并且不需要同时进行蛋白质合成。与在37℃下全程孵育的整个视网膜对照相比,最初在4℃储存会增加尿苷摄取,但不会增加其掺入三氯乙酸可沉淀物质中的量。相反,从4℃转移到37℃后长达72小时内,4℃储存的视网膜中氨基酸的摄取和掺入都会增加。这表明增强的GS活性可能源于4℃储存后总体蛋白质合成增加和特定信使RNA合成增加的共同作用。