Stacey D W, Allfrey V G
Cell. 1976 Dec;9(4 PT 2):725-32. doi: 10.1016/0092-8674(76)90136-7.
Microinjection techniques were used for the study of the translation of heterospecific messenger RNAs in living cells. Duck globin messenger RNAs, in the form of polysomes, messenger ribonucleoprotein particles, or free mRNAs, were injected into cells of human origin (HeLa), and the subsequent appearance and accumulation of avian globin were measured by specific immunofluorescence techniques. Globin accumulated within a high proportion of HeLa cells for at least 25 hr following injection, indicating the intracellular stability of the heterospecific globin messenger RNA and the fact that mRNA initially associated with more complex structures could be translated in the HeLa cytoplasm. In similar experiments, duck reticulocyte polysomes were shown to be translated in duck embryo cells but not in embryonic chick liver cells. The latter, however, could be shown to withstand the microinjection procedure as judged by their capacity to respond normally to inducers and inhibitors of the synthesis of alpha-aminolevulinic acid synthetase. HeLa cells injected with duck hemoglobin divide at normal or nearly normal rates following the injection, but their globin content diminishes, in contrast with the increasing globin content of cells receiving duck reticulocyte polysomes, mRNP particles, or free mRNA.
微注射技术被用于研究异源信使核糖核酸在活细胞中的翻译。以多核糖体、信使核糖核蛋白颗粒或游离信使核糖核酸形式存在的鸭珠蛋白信使核糖核酸被注射到人源细胞(HeLa)中,随后通过特异性免疫荧光技术检测禽类珠蛋白的出现和积累情况。注射后,在很大比例的HeLa细胞中珠蛋白积累至少达25小时,这表明异源珠蛋白信使核糖核酸在细胞内的稳定性,以及最初与更复杂结构相关的信使核糖核酸能够在HeLa细胞质中被翻译这一事实。在类似实验中,鸭网织红细胞多核糖体在鸭胚细胞中可被翻译,但在鸡胚肝细胞中则不能。然而,通过它们对α-氨基乙酰丙酸合成酶合成诱导剂和抑制剂的正常反应能力判断,后者能够耐受微注射过程。注射了鸭血红蛋白的HeLa细胞在注射后以正常或接近正常的速率分裂,但其珠蛋白含量减少,这与接受鸭网织红细胞多核糖体、信使核糖核蛋白颗粒或游离信使核糖核酸的细胞中珠蛋白含量增加形成对比。