McElligott M A, Dice J F
Biochem J. 1983 Dec 15;216(3):559-66. doi: 10.1042/bj2160559.
The technique of erythrocyte-mediated microinjection has been successfully adapted for use with cultured muscle cells. Erythrocytes were fused with primary chick myotube cultures with poly(ethylene glycol), and fluorescent antibodies to haemoglobin demonstrated that this protein was injected into the sarcoplasm of myotubes. The microinjection treatment did not significantly alter protein metabolism in the muscle cells as monitored by rates of synthesis and degradation of muscle proteins. 125I-labelled ribonuclease A and bovine serum albumin were degraded with the expected exponential decay kinetics after microinjection into muscle cells, and the half-life of ribonuclease A (40 h) was approximately twice that of bovine serum albumin (17 h). The degradation of ribonuclease A in the muscle cells was enhanced 1.6-fold in the absence of horse serum and chick-embryo extract, whereas the degradation of bovine serum albumin was not altered during deprivation. These results are characteristic of the breakdown of microinjected ribonuclease A and bovine serum albumin in other cell types. Therefore, our experiments indicate the erythrocyte-mediated microinjection is a valid technique to study protein degradation in primary chick muscle cultures.
红细胞介导的显微注射技术已成功应用于培养的肌肉细胞。用聚乙二醇将红细胞与原代鸡肌管培养物融合，针对血红蛋白的荧光抗体表明该蛋白质被注入到肌管的肌浆中。通过监测肌肉蛋白质的合成和降解速率发现，显微注射处理并未显著改变肌肉细胞中的蛋白质代谢。将125I标记的核糖核酸酶A和牛血清白蛋白显微注射到肌肉细胞后，它们以预期的指数衰减动力学进行降解，核糖核酸酶A的半衰期（40小时）约为牛血清白蛋白（17小时）的两倍。在没有马血清和鸡胚提取物的情况下，肌肉细胞中核糖核酸酶A的降解增强了1.6倍，而在缺乏这些物质的情况下，牛血清白蛋白的降解未发生改变。这些结果是其他细胞类型中显微注射的核糖核酸酶A和牛血清白蛋白分解的特征。因此，我们的实验表明红细胞介导的显微注射是研究原代鸡肌肉培养物中蛋白质降解的有效技术。
